JPH0372485A - Staurosporine derivative - Google Patents

Abstract

NEW MATERIAL:A compound expressed by formula I (R1 and R2 are H, amino, OH or hydroxymethyl, provided that both R1 and R2 are not H). EXAMPLE:3-Aminostaurosporine. USE:A blood platelet agglutination inhibitor. PREPARATION:The methylamino group at the 4'-N-position of a staurosporine expressed by formula II is protected to provide 4'-N-(beta,beta,beta- trichloroethoxycarbonyl)staurosporine, which is then reacted with acetic anhydride to afford an acetyl derivative expressed by formula III (TEC is beta,beta,beta- trichloroethyoxycarbonyl). The resultant derivative is subsequently reacted with nitronium trifluorosulfonate to provide a mononitro derivative, which is then deacetylated to afford a compound expressed by formula IV. The obtained compound expressed by formula IV is subsequently reacted with zinc dust and dilute hydrochloric acid to provide the compound expressed by formula I (group R1 is amino group and group R2 is H).

Description

Detailed Description of the Invention (Industrial Field of Application) The present invention is directed to the following formula ()% formula% (wherein R1 and R2 are each a hydrogen atom, a) group, hydroxy group, which has a platelet aggregation inhibiting effect. , represents a hydroxymethyl group. however,
A staurosporine derivative and an acid addition salt thereof, wherein both R5 and R1 are not hydrogen atoms.

The present invention relates to a staurosporine derivative represented by:

(Prior art) It is already known that staurosporine has a strong vasorelaxant effect and platelet aggregation inhibiting effect (↓).
Japanese Patent Publication No. 53-73501).

(Problems to be Solved by the Invention) If staurosporine becomes highly specific to platelets, it will be clinically useful as a platelet aggregation inhibitor.

(Means for solving problems) The present inventors developed a staurosporine derivative for domestic rabbits.

We searched for the relaxing effect on isolated blood vessels and the inhibitory effect on human platelet aggregation. That is, we conducted intensive research on compounds that have a strong ability to inhibit platelet aggregation and a weak ability to relax blood vessels. As a result, the present invention was completed by synthesizing a staurosporine derivative that has excellent effects with high specificity for platelets.

That is, the present invention represents a group represented by the general formula (I) (wherein R1 and R2 are a hydrogen atom and a hydroxyl group, respectively), a hydroxy group, and a hydroxymethyl group. however,
Both R1 and R2 are not hydrogen atoms.) The present invention relates to a staurosporine derivative and its acid addition salt represented by the formula (R1 and R2 are not hydrogen atoms).

Examples of acids to be added include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, formic acid, acetic acid, benzoic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, citric acid, oxalic acid, methanesulfonic acid, and toluenesulfonic acid. , aspartic acid, glutamic acid, etc.

The compound represented by general formula (I) and its acid addition salt are:
It has a strong platelet aggregation inhibitory effect.

The compound represented by general formula (I) can be produced from staurosporine in several steps. I will explain how to do that.

The methylamino group at the 4°-N-position of staurosporine represented by formula (II) needs to be protected because it has high reaction activity. As the protecting group, a β, β, β-trichloroethoxycarbonyl group (hereinafter abbreviated as TECM) is effective.

In step 1 shown below, the formula (I
Compound II) is synthesized.

(Step 1) The following steps are the same, but the isolation and purification of the product are as follows:
It can be carried out by commonly used methods, such as a combination of extraction, crystallization, chromatography, etc.

A compound of formula (IV) in which an acetyl group is introduced as a protecting group for the amide nitrogen atom at the 6-position can be synthesized by Step 2 shown below.

(Step 2) By using β, β, β-trichloroethyl chloroformate (1.1 to 1.5 equivalents relative to staurosporine) as a reagent, the 4'-N-
(β,β,β-trichloroethoxycarbonyl)staurosporine can be obtained.

The reaction is carried out in a pyridine solvent at temperatures ranging from o'c to room temperature and is complete within a few hours.

The acetyl form of formula (IV) can be obtained by dissolving the compound of formula (III) in 2,6-lutidine and reacting it with acetic anhydride. Usually, 5 equivalents or more of acetic anhydride is suitable for the compound of formula (■).The reaction is carried out at a temperature of 120 to 130°C and is completed within several hours.

Formula (V) and (
The compound Vl) can be synthesized by the following step 3.

(Step 3) Me-N-TEC I can precept.

(Step 4) or Th:C-N-Me A compound of formula (TV) or (III) is reacted with nitronium trifluorosulfonate in dichloromethane to obtain a mononitro compound of formula (V) and a compound of formula (Vl), respectively. A dinitro form can be obtained. Nitronium trifluorosulfonate is 1 for the synthesis of compounds of formula (V).
．． 2-1. 5 equivalents, and for the synthesis of the compound of formula (Vl), it is appropriate to use 10 to 20 equivalents, and the reaction is -60
The test is carried out at temperatures ranging from -78'C to -78'C and is completed within one hour.

Formulas (■) with a substituent (formyl group) introduced into the aromatic ring, (
Compound (■) is synthesized in step 4 by reacting the compound (IV) with titanium tetrachloride and α,α-dichloromethyl methyl ether in dichloromethane to form a monoformyl compound (■) and a diformyl compound (■). Obtainable. Regarding monoform structure synthesis, titanium tetrachloride has 2
~15 equivalents, and α,α-dichloromethyl methyl ether is 1. It is appropriate to use 5 to 3 equivalents, and the reaction is carried out with O
``Carried out at room temperature from C to C and completed in 6 to 12 hours.

In addition, regarding diformyl compound synthesis, titanium tetrachloride has 2
It is appropriate to use 0 to 25 equivalents, and α,α-dichloromethyl methyl ether is used in an amount of 0 to 15 equivalents, and the reaction is carried out within the range of O'C to room temperature and is completed in 6 to 12 hours. .

The synthesis of staurosporine derivatives of formulas (IX) and (X) in which R6 is a hydroxyl group and Rt is hydrogen and hydroxyl group, respectively, can be obtained by the following step 5. In step 5, a Bayer-Villiger type rearrangement reaction is performed followed by deacetylation.

(Step 5) Compounds of formula (■) or (■) are reacted with dichloromethane (■) or (■) with sodium borohydride in an inert solvent to form compounds of formula (X I
) and the dihydroxymethyl form of formula (XI) can be obtained.

Examples of inert solvents include tetrahydrofuran and dioxane. A suitable amount of sodium borohydride is 1.5 to 5 equivalents based on the formyl group of the reaction starting material6.
The reaction is carried out at a temperature ranging from 0°C to room temperature and is completed within a few hours.

Deacetylation of compounds of formulas (V), (XI) and (Xn) is carried out in step 7 as shown below.

A rearrangement reaction was carried out with metachloroperbenzoic acid and potassium hydrogen carbonate in ethanol under the exclusion of light, followed by treatment with an aqueous sodium hydroxide solution, deacetylation, and the respective formulas (
A monohydroxy form of IX) and a dihydroxy form of formula (X) can be obtained. A suitable amount of metachloroperbenzoic acid is 2 to 6 equivalents, and the reaction is carried out at room temperature and completed within several hours.

Formula (X I ) in which R1 is a hydroxymethyl group, and R1 is hydrogen and a hydroxymethyl group, respectively
, (Xrl), the synthesis of staurosporine derivatives is as follows:
Obtained by the next step 6.

(Step 6) (Step 7) A compound of formula (V), (XI) or (Xll) is reacted with hydrazine hydrate in an inert solvent to form (Xll), respectively.
Deacetylated forms of (XrV), (XrV) and (XV) can be obtained. Examples of inert solvents that can be used include methyl cellosolve, tetrahydrofuran, dioxane, etc. Hydrazine hydrate has formulas (V), (XI), and (XII).
The 0 reaction, which is used in large excess with respect to the compound, is carried out at a temperature ranging from 0°C to room temperature and is completed in several hours.

The synthesis of de-TEC staurosporine derivatives of formulas (XVI), (X■), (X■) and (XIX) is obtained by the following step 8. In step 8, a TEC removal reaction is carried out, and at the same time, a nitro group is converted into an amino group.

Formula (XI[I), (Vl), (X), (XV) (7)
The compound is reacted with zinc powder and dilute hydrochloric acid in an inert solvent (Step 8) e-N-H to form a monoamino compound of formula (XVI) and a monoamino compound of formula (X■), respectively.
diamino form of formula (X■), dihydroxy form of formula (X■), formula (XI
A dihydroxymethyl form of X) can be obtained. Furthermore, by this step, corresponding monohydroxy and monohydroxymethyl compounds can be obtained from formulas (IX) and (X, IV), respectively. Examples of inert solvents include methylcellosolve, tetrahydrofuran or dioxane.The reaction is carried out in the range of 0 DEG C. to room temperature and is completed within a few hours.

In addition, the synthesis of a derivative with different substituents introduced at the 3-position and the 9-position, such as 3-amino-9-hydroxystaurosporine (XX), is carried out from a compound of formula (χ■) in the following step. 4. The synthesis can be performed by sequentially performing steps 5 and 8.

(Step 4) (Step 5) compound (Step 8) of formula (XI)
χXI) can be synthesized by sequentially performing Step 3, Step 5 and Step 8 from the compound of formula (■) as follows.

Compound of formula (■) (Step 3) (Step 5) (Step 8) (Action) The staurosporine derivative represented by the general formula (I) of the present invention also has a vasoconstriction inhibitory effect, but has no effect on platelets. It has a more specific and stronger aggregation inhibiting effect on Therefore, thrombosis, in which platelet aggregation is one of the triggers, especially
It is thought to be effective in improving blood flow insufficiency associated with malignant tumors, burns, arteriosclerosis, cerebral infarction, etc.

The staurosporine derivative preparation of the present invention containing the compound represented by the general formula ([) as an active ingredient can be administered orally in the form of a tablet or capsule, or parenterally in the form of a sterile solution or suspension. By prescribing this drug, it is possible to attempt to improve the above symptoms.

Said active ingredient for use in the present invention is administered to patients in need of such treatment in a dosage range of 0.01 to 40 mg per patient, generally in divided doses, thus 0.1 to 200 mg per day. Can be administered in full daily doses.

The dose depends on the severity of the symptoms, the patient's weight, and the person concerned (physicians).
may vary depending on other factors recognized by

Specific drugs that can be mixed into tablets, capsules, etc. are shown below. Binders such as tragacanth, gum arabic, cornstarch, or gelatin; Excipients such as microcrystalline cellulose; Leavening agents such as cornstarch, gelatinized starch, alginic acid, etc.; Lubricants such as magnesium nistearate; Sweeteners such as g-sugar, lactose or saccharin; flavoring agents such as peppermint, redberry oil, or cherry; and when the dosage unit form is a capsule, materials of the above type may additionally contain sweeteners such as fats and oils. A liquid carrier can be included. Various other materials can be present as coatings or to otherwise modify the physical form of the dosage unit.

Sterile compositions for injection include the active substance dissolved or suspended in a vehicle such as water for injection, a naturally occurring vegetable oil such as sesame oil, coconut oil, peanut oil, cottonseed oil, or a synthetic fatty vehicle such as ethyl oleate. It can be formulated according to normal formulation practices. Buffers, preservatives, antioxidants, etc. can also be mixed as necessary.

(Effects of the invention) [Effect on platelet aggregation] Blood collected from a human vein with the addition of 3.8% sodium citrate l/10 volume was centrifuged at 1000 rpm for 10 minutes to prepare platelet-rich plasma (PRP). Next, add 25 μl of phosphate buffer containing PRP200uffi and the test compound to the cuvette of the platelet aggregation needle and mix.
After incubating at 7°C for 3 minutes, add a collagen solution (final concentration 4 μg) as a platelet aggregation inducing substance while stirring.
/m.

3 μg/Idt), adenosine diphosphate (ADP) solution (final concentration 5 μM) or 9.11 to dideoxy 9α, 1
la-methanoepoxy-prostaglandin F2α (U
46619) solution (final concentration 0°3 μM) was added, and changes in permeability due to platelet aggregation were measured. Measurements were performed with various concentrations of the test compound, and the concentration of the compound that inhibited platelet aggregation by 50% using this measurement system, and the IC2° value, were determined. The results are shown in Table 1(^) and Table 1(B).

3-Aminostaurosporine and 3,9-diaminostaurosporine are effective in collagen and ADP aggregation.
Also 3,9-dihydroxystaurosporine and 3.
9-Dihydroxymethylstaurosporine showed a stronger platelet aggregation inhibitory effect than staurosporine in terms of collagen, ADP, and U-46619 aggregation.

Table 1 (A) Compound ICi.

Table 1 (B) [Effect on isolated vascular smooth muscle] Rabbit thoracic aorta Journal Pharmacological Experimental Therapy, Vol. 231, 141-145
It was excised and a vascular strip was prepared as shown on page 1984. In addition, the experimental method also conformed to the conditions described in the above magazine.

The experimental method is outlined below. A vascular strip preparation was suspended isometrically in Krebs-Henseleit solution for 10 d, and a contraction-inducing substance KCI (final concentration 60 mM) was added to perform pre-contraction. After that, wash and add KCt to 10.20, 30,
A cumulative total of 40.60 mM was added to induce contraction, and the tension was used as a drug-free control. Furthermore, a test compound solution was added to the washed blood vessel preparation, and pre-incubation was performed for 1 hour. Thereafter, KCI was cumulatively added (10 to 60 mM), and the contraction inhibition reaction was observed.

The concentration (E D s. value) that inhibits vasoconstriction caused by 40 mM KCI by 50% was determined and shown in Table 2.

3-aminostaurosporine, 3.9-diaminostaurosporine, 3.9-dihydroxystaurosporine and 3.9-dihydroxymethylstaurosporine are:
In KCI contraction, it showed weak contraction inhibition compared to staurosporine.

Table 2 As a result, the compound of the present invention exhibited platelet aggregation effect to the same extent as vasoconstriction or at a lower concentration. In particular, 3.9-cyaminostaurosporine was effective at a low concentration of 0.06 to 0.3 μM. In addition, 3,9-dihydroxystaurosporine causes vasoconstriction of 5. OuM, whereas platelet aggregation inhibition is 1. O~2. Similarly, 3゜9-dihydroxymethylstaurosporine showed vasoconstriction at 0.5 μM and platelet aggregation inhibition at 0.5-1.
.

The effect was shown in luM. On the other hand, it can be seen that staurosporine has a rather stronger effect on vasoconstriction than the compounds of the present invention. That is, this shows that the compound group of the present invention has high specificity for platelets.

(Example) Next, examples will be shown.

Example 1 (i) 93211 g of 4'-N-(β, β, β-trichloroethoxycarbonyl) staurosporin staurosporin (2.0 m5 oz. was dissolved in 1 OIIIi of dry pyridine, cooled to 0°C,
β.

β, β-trichloroethyl chloroformate 0°3dl
(2.2 mmoJ was dropped and reacted for 10 hours.
Add 10 ad of water to the reaction solution! and extracted with chloroform,
The organic layer was dried over anhydrous sodium sulfate.

After filtering off the sodium sulfate, the solvent was removed under reduced pressure, and the residue was purified by column chromatography using silica gel (elution solvent: chloroform) to give pale yellow crystals 4''-N-(
β,β,β-Trichloroethoxycarbonyl)staurosporin 1052■ was obtained (yield 82%).

'HNMR (90MHz, CD CI!
δ) 9.40 (d, IH, J=8 Hz), 8.0
0-7.20 (m, 7H), 6.75-6.68 (m,
IH), 6.62 (br, s, IH), 5.00 (s
, 211), 4.77 (br, s, 2H), 4.07 (
br, s, 111), 2.85 (s, 3H), 2.70
~2.55 (m, IH), 2.60 (s, 3H), 2.
50-2.40 (m, 2H), 2.4Hs, 311) I R (K B r ): 3420, 2950.
1705.1685.1638.1590, 1455.
1395.1383.1345.1310.1283.
1255.1230°1140, 1112.1105.
10?5.1055.1020.810,745,72
0cm-' MS ta/z 640 (M”), 642 (M
"+2), 644(M"+4)(ii) 6
-Acetyl-4°-N-(β,β.

β-Trichloroethoxycarbonyl) Staurosporine 4° -N-(β, β, β-Trichloroethoxycarbonyl) Staurosporine 846 (I, 32 mmojl
was dissolved in 35d of 2,6-lutidine, and dissolved in 12d of acetic anhydride.
d was added dropwise, and the mixture was reacted for 3 hours while heating at 140°C. After adding 40 d of chloroform to the reaction solution, the solution was washed successively with dilute hydrochloric acid, saturated aqueous sodium bicarbonate solution and water, and dried over anhydrous sodium sulfate. After filtering off the sodium sulfate, the solvent was removed under reduced pressure, and the residue was recrystallized from acetone to give pale yellow crystals of 6-acetyl-4°-N-(β).

770 ml of β, β-trichloroethoxycarbonyl) staurosporine was obtained (yield: 85%).

'HNMR (90MHz, CD C13
δ) 9.40 (d, IH, J=8 Hz), 8. OO
~7.20 (m, 7H), 6.73 (m.

1B), 5. OO(s, 2H), 4.80(s, 2H)
, 3.95 (br, s, IH).

2.85 (s, 38), 2.65 (s, 38)
, 2.70-2.55 (n+, 1fl), 2
．． 55 (s, 3H), 2.50-2.40 (m, 2H
), 2.41 (s, 3H) I R (K B r )
: 2930.1?15.1685.1630.159
0,1450,1385.1337.1280.113
5.1110.745cm-'MS w, /z
682 (M"), 684 (M'+2
), 686(M”+4)(ij) 6-acetyl-3-nitro-4°-N-(β,β,β-trichloroethoxycarbonyl)staurosporine Dry dichloromethane 20M1 was cooled to 0°C and trifluoromethane After adding 75 μl of sulfonic acid, add 3 ml of fuming nitric acid.
5 μl was added and stirred for 20 minutes. The reaction solution was cooled to -78°C, and 6-acetyl-4''N-(β,β,β-trichloroethoxycarbonyl)staurosporine 885■(
0.56mm.

40 d of the dichloromethane solution of 1) was added dropwise and the reaction was carried out for 30 minutes.The reaction completed solution was washed with a saturated aqueous sodium bicarbonate solution and water, and dried over anhydrous sodium sulfate.

After filtering off the desiccant, the solvent was removed under reduced pressure and the residue was recrystallized from methanol to give pale yellow crystals of 6-acetyl-3-nitro-4"-N-(β,β,β-trichloroethoxycarbonyl)sulphate. Taurosporin 373■ was obtained (yield 9
1%).

'HNMR (90MHz, CDCli δ) 9.85
(s, III) , 8.00 ~ 6.90 (+++,
611), 6.75 (br, s, IIt), 5.0
0 (s, 211), 4.85 (s, 211), 3.
95 (br, s, 1tl), 2°90 (s, 3+1),
2.70 (s, 6H), 2.40 (s, 3) 1),
2.90~2°40 (m, 311) I R (K B r ): 3400, 2950,
1715, 1695.1600, 1520.1465,
1345,1135,1110.745 cm-'MS
m/z 727 (M"), ?29 (M')
+2), 73BM” +4)(iv) 3-
Nitro-4°-N-(β,β,β-trichloroethoxycarbonyl)staurosporine 6-acetyl-3-nitro-4°-N-(β.

β, β-trichloroethoxycarbonyl) staurosporine 373 (0,51 mmo ff1) was added to methyl cellosolve 50-, followed by hydrazine hydrate (85%) 1
2d was added dropwise, and the reaction was carried out at room temperature for 3 hours. After adding 500 d of water to the reaction-completed solution, it was extracted with chloroform, and the chloroform solution was washed with water and dried over anhydrous sodium sulfate. After filtering off the sodium sulfate, the solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform) to give yellow crystals of 3-nitro-4°-N-(β9β,β-trichloroethoxycarbonyl). ) Staurosporine 274■ was obtained (yield 78
%).

' HN M R (90 MHz, D M S
Od & δ) 9.86 (d, IH, J=2.5 Hz
), 8. OO~7.32 (m, 6H), 7.28 (br
, s, IH), 4,90 (br, s, 4H), 4.0
O(s, IH), 3.30~3,15(*, IH)
, 2.75 (s, 311), 2.90-2.
70 (+s, 11(), 2.53(s, 3H)
, 2.35-2.20 (m, IH), 2.15 (s,
311) I R (K B r ): 3400.2
950.1695.1595, 1520.1465.1
345,1140.1110.745cm-'MS
ts/z 685 (M"), 6B? (M" +2
), [i89(M”+4)(■) 3-aminostaurosporine 3-nitro-4°-N-(β,β,β-trichloroethoxycarbonyl)staurosporine 100g (0
, 146mmo ff1) Methi) Lt-t! T:1'
) ntb 500- and cooled to 0°C, zinc powder 1
゜0g and 1N-hydrochloric acid 10M1 were added sequentially, and 2
After the reaction was completed for 0 hours, 50% of a saturated aqueous sodium bicarbonate solution was added, and insoluble matter such as zinc was filtered off. Chloroform was added to the solution for extraction, and the chloroform solution was washed with water and then dried over anhydrous sodium sulfate. After filtering off the sodium sulfate, the solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform-methanol) to obtain 25 μ of 3-aminostaurosporin as light brown crystals (yield: 36 %).

'HNMR (400MHz, CDCIs δ) 8.7
9 (d, IH, J=2.1 Hz), 7.91 (d
, 18. J=8.5 tLz).

7.86 (d, 18. J=7.511z), 7.3
9 (t, IH, J = 8.0, 8.5 Hz), 7.29 (
t, III, J=7.5, 8.0 Hz), 7.09
(d, ill, J=8.511z), 6.93(dd,
IH, J = 2.1.8.5 Hz), 6.57 (d, I
H.

J=5.7 H2), 5.00(S, 2H), 3.8
4 (d, IH, J = 3.5 Hz).

3.38 (s, 3M), 3.32 (dd, IH, J=3
．． 5,7.1Hz), 2.69 (dd, IH, J=5.
7.16.1 Hz), 2.34 (m, III), 2.
33(s, 3H).

1.54 (s, 38) CNMR (400MHz, CDC136C) 173.
695.139.738.139.546.131.6
91, 131,595.130.794.127,5
37.124.679.124.253, 124.03
0°120.543.119.902.118.412
．． 115.212.115.092゜114.842,
113.851, 111.853.107.42B,
91.077°84.176, 80.141.57.2
5B, 50.353.45.904°33.353.
30.227. 30.045I R(K B r
): 3400,3350.2940.1670
．． 1620. 15B5.1495.148? , 14
55.1320.12B0.1235.1110.75
0 cm' MS ts/z 481 (M") Example 2 (i) 3.9-dinitro-4"-N-(β.

β,β-trichloroethoxycarbonyl) staurosporin 16all dry dichloromethane was cooled to 0°C, 440μl of trifluoromethanesulfonic acid was added, 250μl of fuming nitric acid was added, and the reaction mixture was stirred for 20 minutes.
Cool to 8°C, 4°-N-(β.

A solution of 2001 g (0.31 mmoffi) of β, β-trichloroethoxycarbonyl) staurosporine dissolved in 12 m of dichloromethane was added dropwise, and the reaction was carried out for 45 minutes.The reaction solution was washed with a saturated aqueous sodium bicarbonate solution and water. , dried over anhydrous sodium sulfate. After filtering off the sodium sulfate, the solvent was removed under reduced pressure to obtain yellow crystals of 3,9-dinitro-4'-N-(β,β,β-trichloroethoxycarbonyl)staurosporine 193.
(1) was obtained (yield 85%).

' HN M R (90 MHz, D M S
Od b δ) 9.93 (s, 111), 8.67 (
s, IH), 8.29 (d, IH, J=7.8 Hz)
.

8.20 (d, IH, J = 7.8 Hz), 8.10 (
d, IH, J=8.411z),? ,? ? (d, 111
．． J=8.4 Hz), 7.07 (br, s, 111)
, 5.00 (br.

s, 4H), 4.30 (br, a, IH), 3. lO~
2.10 (+*, 311), 2.73 (s, 311)
, 2.53 (s, 3tl) , 2.40 (
s, 3B) I R (K B r ): 1700
,1595,1518,1465.1340,1302
, 1230.1145.1100.820.797.7
45C! l-'MS■/z 731 (M' +1)
, 733(M"+3), 735(M"+5) (ii) 3.9-Diaminostaurosporine 3.9-dinitro-4°-N-(β,β,β-trichloroethoxycarbonyl)staurosporine 100100a , 137m
moffi) in methyl cellosolve 28IR1,
While cooling to O''C, 2.6 g of zinc powder and 1 IN-hydrochloric acid
5I#1 was sequentially added and the reaction was carried out at room temperature for 2 hours. After the reaction was completed, 60 Jd of a saturated aqueous sodium hydrogen carbonate solution was added, and insoluble materials were filtered off. The solution was extracted with chloroform and dried over anhydrous sodium. After removing the sodium sulfate by filtration, the solvent was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform-methanol) to obtain light brown crystals of 3,9-cya short nostaurosvorin 21.
(3) was obtained (yield 31%).

H-NMR (400MHz, DMSO-d, δ) 8.4
4 (d, IH, J=2.0 Hz), 8.31 (b
r, s, 111), 7.64 (d, LH, J=9.0
Hz), 7.24 (d, IH, J=8.311z), 7
．． 09 (d, IH, J=1.8 Hz), 6.81 (d
d, IH, J = 2.0, 8.3 Hz), 6°74 (d
d, IH, J=1.8.9.0Hz), 6.53(d,
18. J=5.0112).

c, a, 4.80 (br, s, 4fl), 4.79
(s, 2B), 3.98 (br, s.

IH), 3.45 (gi, IH), 3.21 (s, 3H
), 2.48 (m, 11 (), 2.35 (m, IH),
2.21 (s, 3H), 1.63 (s, 311) RoC-
NMR (400MHz, DMSO-d, δC) 172.555, 142.278, 141.482, 1
32.378,131.499゜130.207,13
0.102, 126.858, 124.997, 123
．． 631°118.000.115,303.114.
528.113.677, 113.396°112.8
39,109.379,108.404,104.18
5. 91.07? .

82.671. 80.105. 57.690. 5
0.890. 45.309°40.2B2. 33.
546. 29.685I R (K B r )
: 3330, 2940.1665.160B, 157
0.148B, 1460, 1450.1400, 134
0,1315.12B5,1235.1142゜112
8, 1100.106B, 1040.795.720
cm-'MS (FAB-) 497(M"+
1) Example 3 (i) 6-acetyl-3,9-diformyl-4° −
N-(β,β,β-trichloroethoxycarbonyl)staurosporin 6-acetyl-4°-N in 100 d of dry dichloromethane
-(β,β,β-Trichloroethoxycarbonyl)staurosporin 100g (0,196mmol ff1) was dissolved and cooled to 0°C, titanium tetrachloride 320#j!
(20 eq), further added 130 μl of α,α-dichloromethyl methyl ether (I0 eq), and stirred at room temperature for 2 hours.
The reaction was carried out for 5 hours. Add 10% dichloromethane to the reaction completed solution.
After adding 0d, the mixture was washed with a saturated aqueous sodium bicarbonate solution and water, and dried over anhydrous sodium sulfate. After filtering off the sodium sulfate, the solvent was removed under reduced pressure to give yellow crystals of 6-acetyl-3,9-diformate 4'-N-(
β.

72 mg (0.13 mmof) of β, β-trichloroethoxycarbonyl)staurosporine was obtained (yield 67
%).

' HN M R (90 MHz, D M S
Od & δ) 10, 07 (s, 1ll), 9
．． 83 (s, III), 9.23 (s, IH
), 8.29(s, 1fl), 8.02(
d, IH, J=2.8Hz), 7.93 (d
, IH, J=7.811z), 7°73(d, L
H, J=7.8Hz), 7.60 (d, IH, J=7.
8tlz), 6.91 (br, s, 111), 5.0
6 (s, 2H), 4.99 (s, 2H), 4.26 (b
r, s.

111), 3.10 to 2.10 (m, 311), 2.
73 (s, 3H), 2.53 (s, 3H), 2.52
(s, 3) 1), 2.41 (s, 3H) I R (K B
r): 1720.1695.1640,16
23,1596.13B6, 1337.1295.11
97.1143.1117.807,782,763,
715cm-' MS a/z 738 (M'), 740 (M"
+2), 742 (M" +4) (ii) 3.9-
Dihydroxy-4″-N(β,β,β-trichloroethoxycarbonyl)staurosporine 6-acetyl-3,9-diformyl-4″-N(β,β
, β-trichloroethoxycarbonyl)staurosporine 272■ (0.37 mm.

After adding and dissolving 1) in 50 μl of dichloromethane, 346 μl of metachloroperbenzoic acid and 100 μl of potassium hydrogen carbonate were added.
(2) was added, and the reaction was carried out at room temperature for 3.5 hours while shielding from light. The reaction completed solution was washed with 50 d of saturated aqueous sodium sulfite solution, 50 d of saturated aqueous sodium bicarbonate solution and water,
It was dried with anhydrous sodium sulfate. After removing the desiccant by filtration, the solvent was removed under reduced pressure and the residue was dissolved in methyl cellosolve 45-, and 4N aqueous sodium hydroxide solution 10- was added.
The mixture was stirred at room temperature for 2 hours. The reaction completed solution was diluted with IN-dilute hydrochloric acid 5
After neutralization with 0Id, extraction was performed with dichloromethane, and the dichloromethane solution was washed with water and dried over anhydrous sodium sulfate. After sodium sulfate filtration, the solvent was removed under reduced pressure and the residue was subjected to silica gel column chromatography (
Purification with chloroform-methanol) gave pale yellow crystals of 3°9-dihydroxy-4'-N-(β,β,β-
trichloroethoxycarbonyl) staurosporine 19
6■ was obtained (yield 79%).

'HNMR (90MHZ, DMSO-db+Dzo,
δ) 8.20 (s, IH), 7.78 (d,
IH, J=9.6H2), 7.43(d, IH
, J=9.6Hz), 7.31(s, IN), 7.01
(d, 2H, J=9.611z), 6.83(br, s
, LH), 4.93 (s, 2) 1), 4.91 (s, 2)
) 1), 4.23 (br, s.

IH), 4.0-2.0 (m, 311),
2.73 (s, 3tl), 2.67 (s, 3
fl).

2.24 (s, 311) I R (K B r ): 3350,2900,
1700, 1674.1657.1620, 15B0.
1473.1408.1392.1344.1316.
127? , 1220°1142cm-' MS (FAB-) 689(M" +1) (iii)
3.9-dihydroxystaurosporine 3.9-dihydroxy-4'-N-(β,β.

β-Trichloroethoxycarbonyl) Staurosporin 110 (0,164 mmol) was dissolved in 50 d of methyl cellosolve, and while cooling to 0°C, 7.5 g of zinc powder and 15 d of IN = hydrochloric acid were sequentially added, and the solution was dissolved at room temperature for 2 The reaction was carried out for an hour. After adjusting the pH of the reaction solution to 5 with 0.4N aqueous potassium hydroxide solution, insoluble matter was filtered. The solution was adsorbed on adsorption resin HP-20 (Mitsubishi Kasei), and purified using water-methanol. ), 32 ml of 3,9-dihydroxystaurosporin, which is a light brown crystal, was obtained (yield: 40%).

H-NMR (90MHz, DMSO-d, δ) 9.0 (
s, IH), 8.9Cs, IH), 8.5
(s, III), 8.2(s, III),
7°6 (d, Ill, J=9.4Hz), 7.
2 (d, ill, J=9.4Hz), 7.1
(s, 111), 6.8 (d, 211. J
=9.411z), 6.5 (br, s, III
), 4.8(s, 2H), 4.0(br,s,I
H), 4.0-2.0 (m+3H), 2-8(s, 3H
).

2.3(s, 3H), 2.0(s, 311)r R(K
B r ): 3340, 2900, 1664,
1657, 1620, 1580, 1472.1390.
1346cm-'MS (FAB-) 499(M"
+1) Example 4 (i) 6-acetyl-3,9-di(hydroxymethyl)-4'-N-(β,β,β-trichloroethoxycarbonyl)staurosporine 6-acetyl-3,9-
Diformyl-4°-N-(β,β,β-trichloroethoxycarbonyl)staurosporine 172g (0,2
3mm.

A solution obtained by dissolving 1) in 8 d of dry tetrahydrofuran was added to 2 ml of a suspension of dry tetrahydrofuran containing 49 ml of sodium borohydride, and the reaction was carried out for 2.5 hours at room temperature. 100 ml of water was added to the completed reaction solution. After the addition, extraction was performed with chloroform, and the chloroform solution was washed with water and dried over anhydrous sodium sulfate. After filtering off the sodium sulfate, the solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform-methanol) to give pale yellow crystals of 6-acetyl-3,9-di(hydroxymethyl)- 83.6 μ of 4”-N-(β, β, β-trichloroethoxycarbonyl)staurosporine was obtained (
yield 48%).

' HN M R (90 MHz, D M SO
d hδ) 9.00 (s, II), 7.89
(s, IH), 7.83 (d, In, J=9
．． 0flz), 7°50(d, 111.J=9.0
)1z), 7.46 (d, IH, J=9.0Hz), 7
．． 40(d, IH, J=9.0Hz), 6.93(
m, III), 5.23(s, 2H), 5.13
(s, 211), 4.93(s, 2H),
4.64 (s, 1B), 4.62 (s, l1
l), 4°26 (br, s, IH), 4.0 ~ 3
．． 0 (m, l11), 2.80-2.00 (m, 2H)
, 2.73 (s, 311) , 2.64 (s
, 311), 2.62(s, 3H), 2.
33 (s.

3H) I R (K B r ): 3460.2956,
2891.1?16, 1620.1596, 1460.
1420.1384.1344.1316.1290
cta-'MS (FAB-) 743(M"+
1) (ii) 3.9-di(hydroxymethyl)-4°-
N-(β,β,β-trichloroethoxycarbonyl)staurosporine 6-acetyl-3,9-di(hydroxymethyl)-4°
-N-(β,β,β-trichloroethoxycarbonyl)
After adding Staurosporine 360■ (0°48 mmol) to Methyl Cellosolve 36Ili, hydrazine hydrate (85%) 121I11 was added dropwise, and the reaction was carried out for 30 minutes at room temperature. 100ml of water was added to the completed reaction solution. Thereafter, the mixture was extracted with 100 d of chloroform, and the chloroform solution was washed with water and dried over anhydrous sodium sulfate. After filtering off the sodium sulfate, the solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform-methanol) to give pale yellow crystals of 3,9-di(hydroxymethyl)-4''-N- 254 mg of (β, β, β trichloroethoxycarbonyl)staurosporine was obtained (
yield 75%).

' HN M R (90 MHz, D M SO
d & δ) 9.13 (s, 1B), 8.46
(s, III), 7.82 (s, IH),
7.80 (d, IH.

J=9.0)1z), 7.70-7.20(m, 2H)
, 7.00 (br, s, 18), 5°13 (s, 38)
, 5.0O(s, 2H), 4.67(s, 410), 4
．． 26 (br, s.

11+), 2.70-2.55 (steel, 1tl), 2.7
3 (s, LH), 2.60 (s, 1ll).

2.50-2.40 (m, lH), 2.33 (s, 11
1) I R (K B r ): 34B1,2943
, 1691.167B, 1592, 1455, 1400
．． 1383.133B, 1303.1286.125
5.1230.121? .

1148.1130, 1100.1080 c+a-'
MS (FAB-) 701 (M” +1) (iii
) 3.9-di(hydroxymethyl)staurosporine 3.9-di(hydroxymethyl)-4″-N-(β,β
, β-trichloroethoxycarbonyl)staurosporine 35.81Qg (0.05mm.

1) was dissolved in 15 ml of methyl cellosolve, and while cooling to 0''C, 2 g of zinc powder and 3 d of IN-hydrochloric acid were sequentially added.
The reaction was carried out at room temperature for 5 hours. After the reaction is complete, add 1711j of 25% ammonia water! was added, and insoluble matter such as zinc was removed by filtration. Chloroform was added to the solution for extraction, and the chloroform solution was washed with water and then dried over anhydrous sodium sulfate. After filtering off the sodium sulfate, the solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform-methanol) to give light brown crystals of 3,9-di(
17.8 ml of hydroxymethyl)staurosporine was obtained (yield 66%).

' HN M R (90 MHz, D M SO
d & δ) 9, 13 (s, 1B), 8.40 (
s, IH), 7.82 (d, IH, J=9Hz
), 7.80(s, IH), 7.53(s,
IH), 7.47 (s, 01), 7.40
(d, IL J=911z), 6.67(s,
IH), 5.13(br, s, 2H), 4
．． 93 (s, 2H), 4゜73 (s, 4H
), 4.66(s, IH), 4.06(s,
IH), 4.06(s, Ift).

4.0-3.0 (m, IH), 3.27<s, 3H),
2.5-2.4 (m, 28).

2.30 (s, 311), 1.47 (s, 3H) I R
(KBr): 3370,1666.165
7,1590,1457.1412+ 1396+ 1
350 cta-'MS (FAB-) 525(
M” +1) Example 5 (i) 6-acetyl-9-forman-3-nitro-4
-N-(β9 β, β-trichloroethoxycarbonyl)staurosporine dry dichloromethane 1Il11! 6-acetyl-3-
Nitro-4°-N-(β,β,β-trichloroethoxycarbonyl)staurosporine 110■(0,151m
Dissolve the liquid and add 1 part of titanium tetrachloride while cooling to 0°C.
70 μff1 (I0 eq) and further 67 μ1 (5 eq) of α,α-dichloromethylmethyl ether were added, and the reaction was carried out at room temperature for 25 hours.

After adding 100% dichloromethane to the reaction-completed solution, the mixture was washed with a saturated aqueous sodium bicarbonate solution and water, and dried over anhydrous sodium sulfate. After filtering off the sodium sulfate, the solvent was removed under reduced pressure to give yellow crystals of 6-acetyl-9-
Formyl-3-nitro-4°-N-(β,β,β-trichloroethoxycarbonyl)staurosporine 65■
(0,086 mmoffi) was obtained (yield 57%).

' HN M R (90 MHz, D M S
Od b δ) 9.91 (s, III), 9
．． 88 (s, IH), 8.31 (s, LH
), 8.28 (d, IH.

J=7.8)1z), 8.19 (d, Ill
, J=7.8Hz), 7.76 (d, IH,
J=7°811z), 7.63 (d, IH, J
=7.8Hz), 6.93 (br, s, 1ll
), 5.00(s, 2H), 4.97(s,
211), 4.16 (br, s, III),
3.10~2°10 (m, 311), 2.72
(s, 3H) , 2.57(s, 3H) , 2
．． 51 (s, 311).

2.49 (s, 3H), 2.41 (s, 311) I R
(K B r ): 2900.1?20.169
5,1640,1595,1465,1386.134
1.1295.1197.1143.1110.746
cm-'MS (FD-) 755(Mo) (ii) 9-hydroxy-3-nitro-4°N-
(β,β,β-trichloroethoxycarbonyl)staurosporine 6-acetyl-9-forgolu-3-nitro-4'-N
-(β,β,β-trichloroethoxycarbonyl)staurosporine 264 μ (0.35 mmoj2) was dissolved in 50 μl of dichloromethane, 330 μ of metachloroperbenzoic acid and 100 μ of potassium bicarbonate were added, and under light protection, The reaction was carried out at room temperature for 3.5 hours. The reaction-completed solution was washed with 50 d of a saturated aqueous sodium sulfite solution, 50 d of a saturated aqueous sodium bicarbonate solution, and water, and dried over anhydrous sodium sulfate. After removing the desiccant by filtration, the solvent was removed under reduced pressure and the residue was dissolved in methyl cellosolve 40d.
10 ml of 4N aqueous sodium hydroxide solution was added, and the mixture was stirred at room temperature for 2 hours. After the reaction completed solution was neutralized with 50 d of IN-hydrochloric acid, it was extracted with dichloromethane, and the dichloromethane solution was washed with water and dried over anhydrous sodium sulfate. After filtering off the sodium sulfate, the solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform-methanol) to give yellow crystals of 9-hydroxy-3-nitro-4'-N-(β, β.

β-trichloroethoxycarbonyl) staurosporine l 71+ag (0,24 mmof) was obtained (69
%).

' HN M R (90 MHz, D M S
Od & δ) 9.91 (s, 1) 1) , 8.
90 (s, IH), 8.28 (d, ill
, J=7.8Hz), 8゜19(d, ill, J
=7.811z), 7.60(d, LH, J=7.8H
z), 7.16(s, l1l), 6.83(d,
LH, J=7.8Hz), 6.71 (br,
! ! , 1) 1) , 5°0f (s, 111), 4.
96 (s, 111), 4.18 (br, s, 11), 3
．． 10-2.10 (m, 311), 2.73 (s, 3H
), 2.51 (s, 3H), 2.40 (s, 311) I R (K B r ): 340B, 2950.1
? 20.1695.1632.1590, 1455.1
386.1295.1196.1142.1116c
m-'MS (FD-) 701(M') (iii) 3-amino-9-hydroxystaurosporine 9-hydroxy-3-nitro-4'-N-(β.

120 trg (0,171 mmol) of β, β-trichloroethoxycarponyl) staurosporin was dissolved in 500 d of methyl cellosolve, 1.0 g of zinc powder and 1 OId of IN-hydrochloric acid were sequentially added under cooling in an O''C6 oven, and the solution was dissolved at room temperature for 2 hours. After the reaction was completed, 50% of a saturated aqueous solution of sodium hydrogen carbonate was added, and insoluble substances such as zinc were filtered off.Chloroform was added to the solution for extraction, and the chloroform solution was washed with water. Dry with sodium sulfate. After filtering off 1N of sodium sulfate, the solvent was removed under reduced pressure.
The residue was purified by silica gel column chromatography (chloroform methanol) to obtain light brown crystals of 3Aξ.
27 ml of no-9-hydroxystaurosporine was obtained (yield 32%).

' HN M R (90 MHz, D M S
Od b δ) 8, 70 (s, 18), 8.44
(s, IH), 8.31 (br, s, 1B
), 7.58 (d.

lH, J=9.4Hz>, 7.23 (s, 1
B), 7, 10 (d, III, J=8.5
11z).

6.82 (d, LH, J=9.4Hz), 6.
81 (d, 1fL J=8.511z), 6.
53 (br, s, 18), 4.78 (s, 28), 5.
0~4-5(br,s+211)+3°97(br,s
, IH), 3.50-2.10 (m, 3H), 3.16
(s, 311).

2.16 (s, 311), 1.69 (s, 3H) I R
(KBr): 3330.2940.166
B, 1601.1570.148B, 1460.14
52.1403C11-'MS (FAB-) 498
(M” +1) Example 6 (i) 6-acetyl-9-hydroxymethyl-3-
Nitro-4°-N-(β,β,β-trichloroethoxycarbonyl)staurosporine 6-acetyl-9-formyl-3-nitro4′-N-(β,β,β-trichloroethoxycarbonyl)stauros Porin 160■ (
A solution of 0,211 mmof) dissolved in 8 d of dry tetrahydrofuran was added to 2 d of a suspension of dry tetrahydrofuran containing 47 μm of sodium borohydride, and 2 d of dry tetrahydrofuran was added at room temperature.
．． The reaction was carried out for 5 hours. After adding 100 d of water to the reaction-completed solution, it was extracted with chloroform, and the chloroform solution was washed with water and dried over anhydrous sodium sulfate. After filtering off the sodium sulfate, the solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform-methanol) to give yellow crystals of 6-acetyl-9hydroxymethyl-3-nitro-4"- N-(β.

76 ml of β, β-trichloroethoxycarbonyl) staurosporin were obtained (47%).

'H-NMR (90MHz, DMSO-d, δ) 9.9
0 (s, LH), 8.25 (d, ltl,
J=8.5Hz), 8.16 (d, IH,J=
8.5Hz), 7.86 (s, IH), 7.51 (d,
IH, J=9.0Hz), 7.38(d, Ill,
J=9.0Hz), 6.84 (m, l1l)
, 5.03 (s+2H) , 4.89(s,
2H), 4.81 (s, 211), 4.6
3 (s, 18), 4.21 (br, s, IH
).

4.0-2.3 (m, 3H), 2.76 (s, 3H),
2.61 (s, 311), 2.54 (s, 311)
, 2.30 (s, 3H)I R(K B r
): 2951, 2890, 1715, 1620
,1596,1520,1460.1420.13B3
．． 1347.1320.1300 ctn-'MS
(FD-) 757 (drunk) (ii) 9-hydroxymethyl-3-nitro 4°
-N-(β,β,β-trichloroethoxycarbonyl)
Staurosporine 6-acetyl-9-hydroxymethyl-3-nitro-4
"-N-(β,β,β-trichloroethoxy'carbonyl)staurosporine 348mg (0°459mmof
) to methyl cellosol 35-? After being dissolved, 12 mj of hydrazine hydrate (85%)! 3 drops at room temperature.
The reaction was carried out for 0 minutes. After adding 100% of water to the reaction-completed solution, extraction was performed with 100IR1 of chloroform, and the chloroform solution was washed with water and dried over anhydrous sodium sulfate. After filtering off the sodium sulfate, the solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform-methanol) to give yellow crystals of 9hydroxymethyl-3-nitro-4°-N-(β).

β,β-trichloroethoxycarbonyl)staurosporin 221■ was obtained (yield 67%).

' HN M R (90 MHz, D M S
Od h δ) 9, 91 (s, 1) 1) , 8.
40 (s, 1)I), 8.22 (d, In
, J=8.511z), 8°16(cl, 11
1. J=8.5H2), 7.88 (s, IH
), 7.50 (d, IH, J=9°Hlz)
, 7.36 (d, IH, J=9.1llz) ,
6.78 (s, IH), 5.02 (s.

21υ, 4.91 (s, 211), 4.86 (s, 2H
), 4.61 (s, 111), 4.20 (br, s,
IH), 4.10-2. IO(m, 311),
2.75(s, 3H), 2.54(s, 3)1
), 2.31 (s, 3) 1) Summer R (K B r )
: 3430, 2944, 1690, 1677.1
592, 1544, 145B, 1400.1383.
1338.1302cm-'MS (FD-) 715
(M”) (Stone) 3-Awano 9-Hydroxymethylstaurosporine 9-Hydroxy-3-nitro-4°-N-(β.

β,β-trichloroethoxycarbonyl) staurosporine 35aig (0,049 mmol) was dissolved in methyl cellosolve 15111, and 2 g of zinc powder and IN-hydrochloric acid 3I11 were sequentially added to O'C under cooling, and the solution was dissolved at room temperature for 5
A time reaction was performed. After the reaction is complete, add 1 part of 25% ammonia water.
7d was added, and insoluble matter such as zinc was filtered off. Chloroform was added to the solution for extraction, and the chloroform solution was washed with water and then dried over anhydrous sodium sulfate. After filtering off the sodium sulfate, the solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform-methanol) to obtain light brown crystals of 3-amino-9-hydroxymethylstaurosporin 14. (Yield 56%).

' HN M R (90 MHz, D M S
Od b δ) 8, 44 (s, IH), 8.4
1 (s, III), 7.86 (s, LH)
, 7.47 (d, Ill.

Claims (1)

[Claims] General formula (I) ▲Mathematical formulas, chemical formulas, tables, etc.▼(I) (In the formula, R_1 and R_2 represent a hydrogen atom, an amino group, a hydroxy group, and a hydroxymethyl group, respectively.However, R_1 and R_2 cannot both be hydrogen atoms.) A staurosporine derivative and an acid addition salt thereof.