JPH03220194A - Staurosporine carboxylic acid derivative - Google Patents

Abstract

NEW MATERIAL:The compound of formula I (R and R' are H, carboxyl, etc., provided that both R and R' are not H at the same time) and its salt. EXAMPLE:3-Carboxylstaurosporine. USE:A blood platelet aggregation inhibiting agent useful for preventing relapse of thrombosis. It has weak vasoconstriction suppressing action and low toxicity. PREPARATION:The objective compound can be produced by reacting staurosporine of formula II with beta,beta,beta-trichloroethyl chloroformate to protect the 4'-N-site, dissolving the product in pyridine, reacting with acetic anhydride, reacting the resultant acetylated product of formula III (TEC is beta,beta,beta- trichloroethoxycarbonyl) with TiCl4 and alpha,alpha-dichloromethyl methyl ether to obtain the compound of formula IV, reacting the compound successively with an aqueous solution of potassium permanganate and an aqueous solution of NaOH, deacetylating the reaction product and finally reacting the product with zinc powder and dilute hydrochloric acid to remove the TEC group.

Description

Detailed Description of the Invention (Industrial Application Field) The present invention is directed to the following formula (1), which has a platelet aggregation inhibiting effect: or represents an alkoxycarbonyl group that may be divided into two groups, and R and R1 may be the same or different. However, R and R1 are not both hydrogen.

) and its salts.

Furthermore, the following formula (b) (wherein R2 and R3
represents hydrogen, a formyl group, or a carboxyl group, R4 represents hydrogen or an R'CO group, and R5 represents an alkyl group having 1 to 3 carbon atoms.

) The present invention relates to a staurosporine derivative represented by

(Prior Art) It is already known that staurosporine has a strong blood vessel relaxing effect and a platelet aggregation inhibiting effect (Japanese Patent Publication No. 53076/1983 and Japanese Patent Application Laid-open No. 2-69819).

Staurosporine has both a strong platelet aggregation inhibitory effect and a vasoconstriction inhibitory effect, so its efficacy is not specific and it is highly toxic. Platelet aggregation inhibitors that are highly specific to platelets and highly safe are clinically useful, and there is a need to develop such platelet aggregation inhibitors.

(Usage to be Solved by the Invention) (Means for Solving the Problems) The present invention contemplates that staurosporine derivatives have an inhibitory effect on the contraction of isolated blood vessels in rabbits, an aggregation inhibitory effect in guinea pig platelets, and We searched for acute toxicity. That is, we conducted intensive research on compounds that have a strong ability to inhibit platelet aggregation, a weak ability to suppress vasoconstriction, and a weak toxicity. As a result, we succeeded in synthesizing a staurosporine derivative that has excellent effects that are highly specific to platelets and is also low in toxicity.
The invention has been completed.

That is, the present invention is based on the following formula: However, both R and R1 may not be hydrogen.

) and its salts.

In the case of acid addition salts, examples of the acid to be added include hydrochloric acid,
Examples include hydrobromic acid, sulfuric acid, nitric acid, I acid, acetic acid, benzoic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, citric acid, oxalic acid, methanesulfonic acid, toluenesulfonic acid, aspartic acid, and glutamic acid. Also, R and R1
Carboxyl staurosporine, in which at least one of the groups is a carboxyl group, easily forms a carboxylate salt.

Carboxylic acid salts include alkali metal salts, alkaline earth metal salts, and ammonia and amine addition salts.

Alkali metal salts include lithium salts, sodium salts, potassium salts, etc.Alkaline earth metal salts include
There are calcium salts, barium salts, etc., and amine addition salts such as pyridinium salts, trimethylamine salts, triethylamine salts, dimethylamine salts, diethylamine salts, etc.

The compound represented by the general formula ([) and its salts have low toxicity and specificity for the harmful effect of platelet aggregation 'JltII.

The compound represented by the general formula (+) can be easily and efficiently produced from staurosporine via the intermediate represented by the general formula (n). I will explain how to do that.

The methylamino group at the 4'-N-position of staurosporin has high reactivity and needs to be protected. As the protecting group, a β, β, β-trichloroethoxycarbonyl group (hereinafter abbreviated as TBG group) is effective.

The formula ([
Compound [l) is synthesized.

The 4°-N-
(β,β,β-trichloroethoxycarbonyl)staurosporine can be obtained. The reaction is carried out in a halogenated hydrocarbon Xi3 medium such as chloroform in the presence of pyridine, lutidine, or triethylamine as a base, preferably triethylamine, at a temperature of -10 to 50°C, preferably 0°C to room temperature, and has a temperature of 1 to 48°C. hours, preferably 12-2
It will be completed within 4 hours. The same goes for the following steps,
Isolation and purification of the product can be carried out using commonly used methods, such as
This can be carried out by combining extraction, crystallization, chromatography, etc. Further, the reaction solvent in the present invention is the same in the following steps, but a solvent inert to the reaction or a mixture thereof can be used.

A compound of formula (rV) 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.

The compound of formula (III) is pyridine, 2,6-lutidine,
Preferably dissolved in pyridine and reacted with acetic anhydride,
An acetyl form of formula (IV) can be obtained. Usually, 5 equivalents or more of acetic anhydride is suitable for the compound of formula (fir).
It is carried out at a temperature of 120°C and is completed within a few hours.

Compounds of formulas (V) and (Vl) in which a substituent (formyl group) was introduced into the aromatic ring were synthesized by the following step 3.

A compound of formula (IV) is reacted with titanium tetrachloride and α,α-dichloromethyl methyl ether in a halogenated hydrocarbon such as dichloromethane chloroform, preferably in dichloromethane to form the form (Step 4) of formula (V), ( The hydroxylated compound can be obtained by reacting the compound Ml) with an aqueous potassium permanganate solution in an inert solvent. For monoformylation, titanium tetrachloride is 2 to 30 equivalents, preferably IO to 15 equivalents, and also α
, α-dichloromethyl methyl ether is suitably used in an amount of 1 to 4 equivalents, preferably 1 to 2 equivalents, and the reaction is carried out in an amount of 1 to 20 equivalents.
~5℃, preferably within the range of -10~0℃, 6
It will be completed in ~48 hours. In addition, regarding diformylation, titanium tetrachloride is used in an amount of 5 to 25 equivalents, preferably 10 to 20 equivalents.
It is appropriate to use 5 to 15 equivalents of α,α-dichloromethyl methyl ether, preferably 10 to 15 equivalents, and the reaction is carried out in the range of 0°C to room temperature, and 6 to 1
It will be finished in 2 hours.

The compounds represented by the formulas (■) and (■) obtained by converting the formyl group into a carboxyl group and deacetylating them can be obtained by the following step 4.
is executed.

It is reacted with an aqueous solution of thorium, potassium hydroxide, etc., preferably an aqueous sodium hydroxide solution, to deacetylate the formula (
The carboxylic acid forms shown by (■) and (■) can be obtained. Examples of inert solvents that can be used include methyl cellosolve, tetrahydrofuran, dioxane, etc., preferably dioxane.

Regarding the oxidation of the monoformyl compound, it is appropriate to use 1 to 4 equivalents of potassium permanganate, preferably 1 to 2 equivalents, and the reaction is carried out within the range of 0°C to room temperature and is completed in 2 to 12 hours. . Regarding deacetylation, the appropriate amount of sodium hydroxide is 1 to 5 equivalents, preferably 1 to 3 equivalents, and the reaction is carried out at a temperature of 0°C to room temperature and is completed within 1 hour.

Regarding the oxidation of the diformyl compound, it is appropriate to use 2 to 6 equivalents of potassium permanganate, preferably 2 to 3 equivalents, and the reaction is carried out within the range of 0°C to room temperature and is completed in 2 to 12 hours. . Regarding deacetylation, the appropriate amount of sodium hydroxide is 1 to 5 equivalents, preferably 1 to 3 equivalents, and the reaction is carried out at a temperature of 0°C to room temperature and is completed within 1 hour.

Synthesis of the TEC-depleted carboxyl staurosporin represented by the formula (DOl(X)) is carried out by the following step 5.

(Step 6) A methyl ester of formula (XI) can be obtained by reacting the compound of formula (X) with diazomethane in an inert solvent. Examples of inert solvents include methyl cellosolve, tetrahydrofuran or dioxane, preferably methyl cellosolve. Diazomethane is 1-100
It is appropriate to use an equivalent amount, preferably 1 to 20 equivalents, and the reaction is carried out at a temperature ranging from 0° C. to room temperature and is completed in 5 to 10 minutes.

(Function) Staurosbo formula (■) represented by the general formula (r) of the present invention
, (■) can be reacted with zinc powder and dilute hydrochloric acid in an inert solvent to obtain carboxyl staurosporins of formulas (IX) and (X). Examples of the inert solvent include methyl cellosolve, tetrahydrofuran, and dioxane, with methyl cellosolve being preferred. The reaction is carried out at a temperature ranging from 0° C. to room temperature and is completed within a few hours.

Furthermore, in the compound represented by formula (1), R and R
When 1 is a straight or branched alkoxycarbonyl group having 5 or less carbon atoms, it can be easily converted from a carboxyl group. That is, it is carried out by dehydration condensation with alcohol, substitution reaction with alkyl halide, and further esterification reaction with diazoalkane. Among these, a method using a diazoalkane is preferred in consideration of reaction yield and ease of treatment.

For example, the methyl ester compound can be synthesized by Step 6 shown below.

Phosphorus derivatives also have a vasoconstriction inhibitory effect, but have a more specific aggregation inhibitory effect on platelets. Therefore, it is considered to be effective in improving thrombosis caused by platelet aggregation, especially blood flow failure associated with malignant tumors, burns, arteriosclerosis, cerebral infarction, etc.

The staurosporine derivative preparation containing the compound represented by the general formula (1) of the present invention as an active ingredient can be administered orally in a preparation such as a tablet or capsule, or parenterally in a sterile solution or suspension. The above symptoms can be improved by prescribing the drug.

Said active ingredient for use in the present invention is generally administered in doses ranging from 0.01 to 40 μ per patient in several divided doses for patients in need of such treatment, thus 0.1 to 200 per day. ■Can be administered at a total daily dose of .

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

Specific drugs that can be mixed into tablets, capsules, etc. are shown below. tragacanth, gum arabic,
binders such as corn starch or gelatin; excipients such as microcrystalline cellulose; leavening agents such as corn starch, gelatinized starch, alginic acid, etc.; lubricants such as magnesium nistearate; sia sugar, lactose or saccharin. sweetening agents such as peppermint, red radish oil or cherry;1 and when the dosage unit form is a capsule, the above type of material may further contain a liquid carrier such as an oil or fat. I can do it.

Various other materials can be present as coatings or to otherwise modify the physical form of the dosage unit.

Sterile compositions for injection include dissolving the active substance 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. or suspension, according to conventional formulation practices. &l Shocking agents, preservatives, antioxidants, etc. can be mixed as necessary.

(Effect of the invention) [Effect on platelet aggregation] 3.8% sodium citrate 1/l from guinea pig vein
The blood collected by adding O volume was centrifuged at 1000 rpm for 10 minutes to prepare platelet rich plasma (PRP). Next, 25 μl of phosphate buffer containing 200 μe of PRP and the test compound were added to the cuvette of a platelet aggregometer and mixed.
After incubating at 37°C for 3 minutes, a collagen solution (final concentration 5 μg) was added as a platelet aggregation inducing substance while stirring.
/IRIl), adenosine diphosphate (ADP) solution (final concentration 4 μM) and 9,11-dioxy-9α, 11a
~ Methanoepoxy-prostaglandin F2α (U46
619) solution (final concentration 0.5 μM>25 μl was added and the change in permeability due to platelet aggregation was measured. Measurements were performed with various concentrations of the test compound, and this measurement system inhibited platelet aggregation by 50%. The concentration of the compound, IC, was determined.

3-carboxyl staurosporine (IX), 3°9
Table 1 shows the results of the platelet aggregation inhibitory effects of dicarboxyl staurosporine (X) and staurosporine on collagen, ADP, and U466I9 aggregation.

Table 1 [Effects on isolated vascular smooth muscle] Rabbit thoracic aorta with Digital Pharmacological Experimental Terra - Vol. 231, 141-145
(1984), and a vascular strip was prepared. Also, the experimental method meets the conditions described in the magazine above! 1! did.

An outline of the experimental method is given below. The vascular strip preparation was suspended isometrically in 10 mN Krebs-Henseleit solution, and the tension was induced by a contraction-inducing substance KCI (final concentration 60 mM), and the tension was used as a drug-free control. Furthermore, a test compound solution was added to the washed blood vessel preparation and preincubated for 1 hour. Thereafter, KCI was cumulatively added (10 to 60 mM), and the contraction inhibition reaction was observed.

The concentration (EDS. value) that inhibited vasoconstriction caused by 40 mM KCI by 50% was determined, and the results are shown in Table 2. Staurosporine derivatives exhibited weak contraction inhibition of 1/150 to 1/2000 compared to staurosporine in KCI contraction.

Table 2 ICs of platelet aggregation inhibition (U46619). Value and ED of vasoconstriction inhibition. The values were obtained by comparing the values and are shown in Table 3.

Table 3 was prepared, and the LDso value was determined from the amount of the test compound which was intraperitoneally administered once to 5-week-old male Slc:ICR mice, and the mortality rate of the mice was 50%. The results are shown in Table 4. 3゜9
- Dicarboxylstaurosporin LD compared to Rosporin. The value was large and the toxicity was reduced.

Table 4 As a result, the compound of the present invention has a weaker action on blood vessels than staurosporine, and tends to be platelet-specific. That is, the compounds of the present invention are expected to be clinically effective in preventing thrombus recurrence due to their vasoconstriction-inhibiting action and low blood pressure lowering action.

[Acute toxicity in mice]

The test compound was added to a 0.5% sodium carboxymethylcellulose solution to prepare suspensions of various concentrations.As a result, the compound of the present invention has a significantly reduced vasoconstriction inhibitory effect and toxicity compared to staurosporine, and has a significant effect on platelets. The aggregation inhibition effect became more specific. In other words, the compounds of the present invention can be said to be platelet aggregation inhibitors that are highly specific to platelets and highly safe.

(Example) Next, examples will be shown.

Example 1 (i>4'-N-(β,β.β-trichloroethoxycarbonyl)staurosporin Staurosporine 93
2 (2.0 mmof) was dissolved in 10 ml of dry pyridine, and while cooling to 0°C, 0.3 ml (-2.2 mmof) of β1 β,β-trichloroethyl chloroformate was added dropwise and reacted for 10 hours. .10 mj of water in the reaction solution!
was added, extracted with chloroform, and 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 (exiting solvent: chloroform) to give pale yellow crystals 4' -N- (β, β, β- (trichloroethoxycarbonyl) staurosporine 1052■ was obtained (yield 82%).

' H NMR (90 MHz CDCh
δ): 9, 40 (d, IH, J=8 tlz), 8.
00-7.20 (m, 78), 6.75-6.68 (m
, IH), 6.62 (br.s, Ill), 5.00
(s, 211), 4.

77 (br.s, 2H), 4.07 (br.s.
s, IH), 2.85 (s, 3H), 2
．． 70-2.55 (m IH), 2.60 (s, 3)
1), 2.50-2.40 (m.2) 1).

2,41(s,3H) I R (K B r ): 3420,29
50.1705, 1685, 1638.1590, 1
455, 1395, 1383. 1345, 13
10, 1283, 1255, 1230, 114
0, 1112, 1105. 10?5, 1055
, 1020,810,745,720 ω1 MS m/z :640 (M”), 64
2(M"+2), 644(M"+4) (ii) 6-acetyl-4°-N-(β,β.

β-Trichloroethoxycarbonyl) Staurosporine 4° -N-(β,β,β-Trichloroethoxycarbonyl) Staurosporine 846■ (1.32 mmo
1) was dissolved in 2,6-lutidine 35-, and acetic anhydride 1
2- was added dropwise, and the mixture was reacted for 3 hours under heating at 140°C.

After adding chloroform 40- 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'-11-(β
.

β,β-Trichloroethoxycarbonylrosporin 7
70 mg was obtained (yield 85%).

' H N M R (90 MIIZ, CDCl
2 6): 9, 40 (d, J=8 tlz), 8.
00 ~'1.20 (m, 711), 6.73 (m.

111), 5.00 (s, 211), 4.80 (s,
2H), 3.95 (br.s, ltl), 2.85 (
s. 3tl), 2.65 (s. 38), 2
．． 70-2.55 (m, ill), 2.

55 (s, 311), 2.50 ~ 2.40 (m.2
11), 2.41 (s, 311) + R (K B
r): 2930,1715.1685,163
0,1590,1450, 1385cm-' MS m/z: 682 (M”), 684
(M'' +2), 686 (M' +4) (iii) 3-formyl-6-acetyl-4'N~
(β.β,β-trichloroethoxycarbonyl)staurosporine 6-acetyl 4-
rl-(β,β,β-trichloroethoxycarbonyl)
Staurosporine 1. 0g (1.4 mmo
f) and cooled to -1.0°C, titanium tetrachloride 1
．． 5mffi (1 3.6eq), further α
．． α-dichloromethyl methyl ether 0.3d (2.
2 eq) was added thereto, and the reaction was carried out at -10°C for 13 hours. After adding 250 g of 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, and the residue was purified by column chromatography using silica gel (eluent: chloroform-hexane) to give pale yellow crystals of 3-formyl-6-acetyl-
4”N-(β.β,β-trichloroethoxycarbonyl)staurosporine 50 7mg (0.71
mm.

1) was obtained (yield 51%).

' H N M R (90 MHz, CDCl2
6): 9, 90 (s, 1N), 9.33 (s, I
ll), 8.20 ~ 7-20 (m, 611) + 7-0
3 (br.s, IH), 5. 10(s, Il
l), 5.00 (s, 21(), 4.27
(br.

s, 1N) , 2.77 (s, 311) , 2.53
(s, 3H) , 2.40 (s, 311) , 3, 70
〜2,00(m,311)IR(K
B r ): 3483,2947.1718.
1683,1636.1589cm-'MS m/z: 7]0 (M'), 71
2(M"+2), 714(M"+4) (iv) 3-carboxyl-4'-N-(
β.

β,β-trichloroethoxycarbonyl)staurosporine 3-formyl-6-acetyl-4°-N-(β.

β,β-trichloroethoxycarbonyl)staurosporine 850a+g (1.19 mmo6)
After dissolving in 60 ml of 1,4 dioxane, 0.12
15 mf of M potassium permanganate aqueous solution was added, and the mixture was allowed to react at room temperature for 5 hours. After the reaction is complete, IN-hydrochloric acid is added and p
H-3 was prepared to obtain crude crystals. The crude crystals were dissolved in 20% of methyl cellosolve, 2.5% of an aqueous solution of IN=sulfur hydroxide was added, and the reaction was carried out at room temperature for 20 minutes.

Thereafter, IN hydrochloric acid was added to the reaction solution to adjust the pH to -3, and 3-carboxyl-4'-N-(β,β,β-trichloroethoxycarbonyl)staurosporine was precipitated.

The crude crystals were purified by silica gel column chromatography (elution solvent: chloroform-methanol) (yield: 40%).

' H N M R (400 MHz, DMSO
-da δ): 12, 65 (br.s, LH), 9
．． 97(d, 111, J=1.Ol(z), 8.69(
s, IH), 8.09 (dd, ltl, J=1.
0,8.6Hz), 8.08(d, Ill, J=7
．． 8Hz), 8.00 (d, III, J=8.6Hz)
, 7.72 (d, IH.J=8, 6) 1z) , 7.
52 (d, IH, J=8.6Hz), 7.38
(t. J=7.8)1z), 7.10(t, II
(, J=7.5Hz), 5.03(s, 2H)
, 5.02 (s, 211).

4, 66 (d, IH, J=13Hz), 4.3
6 (d, IH. J=16Hz), 2.82 (m
, 11), 2.76 (s, 311), 2.35 (s, 6
8), 2.29 (m, III) I R ( K B r
): 3400, 2995, 1716.16B4
,1592,1469, 1452, 1406, 1
352. 1306, 1222. 1148 cm
- 'MS m/z: 685 (M' +
1)(v) 3-carboxyl staurosporine 3-carboxyl-4'-N- (β,β,β-trichloroethoxycarbonyl)staurosporine 9
30 mg (1.36 mmol) was dissolved in 200 ml of methyl cellosolve, 120 g of zinc powder and 45% of 5N hydrochloric acid were added, and the reaction was carried out at room temperature for 10 minutes. After the reaction is complete, insoluble materials are filtered off and distilled water is added to 200 mL of water.
was added, and the aqueous solution was washed with chloroform.

The 1'8 solution was allowed to stand at room temperature to precipitate pale yellow crystals of 3-carboxyl staurosporine. The crystals were obtained by filtration (yield 69%).

t(N M R(400Ml!z, DMSOJb
δ): 12.66 (br, s, Ift), 9.99 (
d, IH, J=1.611z), 8.89 (br.

s, Ift), 8.71 (s, Iff),
8.08-8.13 (m, 3tl), 7.65
(dit!, J=8.4Hz), 7.57 (d
d, ill, J=8.4.7.3tlz), 1
．． 42 (t, III, J=7.3Hz), 7
．． 01 (dd, IH, J=3.0.9.611z)
, 5゜02(s, 211) , 4.47(s,
Ift), 4.07 (br, s, IH),
3.32 (m, IH), 2.70 (s, 3H)
, 2.50 (s, 3tl) , 2.27 (s
, 3tl), 2°11 (dt, III, J=3.
0,13.011z)' 3CN M R (400Ml
lz,' DMSO-d6171.647. 168.
187. 138.504゜133.178. 129
．． 728. 128.485°126.608. 12
5.833. 123.838°], 22.375 1
21.998 120.965δ) 137.933° 126.979° 122.436° 119.882° 115.180. 114.873. 112.789
．． 109.006°93.368. 81.083
．． 79.272. 60.063゜53.414
．． 45.489. 30.646. 27.7
79゜26.983 I R (K B r ): 3500,3360,
1718, 1640, 1593, 1469, +452
．． 1418,1406.1352.1300.1211
．． 1168cm' MS mHz: 511 (M" +4
) Example 2 (i) 6-acetyl-3,9-diformyl 4' -
N-(β,β,β-trichloroethoxycarbonyl)staurosporine 6-acetyl-4゛N-(β
, β, β-trichloroethoxycarbonyl) staurosporine 100 μm (0,196 mmof) was dissolved,
Titanium tetrachloride 320μN (20eq) under cooling to ℃
, and further α,α-dichloromethyl methyl ether 130
μf (loeq) was added and the reaction was carried out at room temperature for 25 hours. After adding 100 ml of 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 removing sodium sulfate by filtration,
By removing the solvent under reduced pressure, 72 mg of yellow crystal 6-acetyl-39-diformyl-4'-N-(ββ,β-trichloroethoxycarbonyl)staurosporine (
0.13 mmo#) was obtained (67% yield).

tl N M R (90 MHz, CDCh δ
): 10.07 (s, l1l), 9.83 (s, 11
1), 9.23 (s, IH), 8.29 (sIH) 8
．． 02 (d, ltl, J=2.8Hz), 7.93 (d
, III, J=7.8Hz), 7.73(d, IH, J
=7.8)1z),7.60(d,IH,J=7.81
1z), 6.91 (br, s, IH), 5.06
(s, 2H) , 4.99 (s, 28) , 4.26 (
br, s, 1) (), 3.10-2.10 (m, 3H)
, 2.73 (s, 3H), 2.53 (s 3) 1), 2
．． 52 (s, 38), 2.41 (s, 311) I R (
KBr): 1720, 1695, 1640
, 1623.1596cm-'MS mHz:
738 (M”), 740 (M”+2), 74
2(M”+4) (ii) 3.9-dicarboxyl-4°-N(β, β
, β-trichloroethoxycarbonyl) staurosporine 6-acetyl-3,9-diformyl-4゛-N (β, β
, β-trichloroethoxycarbonyl)staurosporine 906 (1,22 mm.

After dissolving 1) in 1,4-dioxane 100-, 2
．． 20 ml of 5mM potassium permanganate aqueous solution was added, and the reaction was carried out at room temperature for 1.5 hours. After the reaction is complete, water 2
After adding 00-, the pH was adjusted to -3 with 5N hydrochloric acid to precipitate crude crystals. The crude crystals taken out by filtration were dissolved in 40 ml of methyl cellosolve, 3 all of an IN-sodium hydroxide aqueous solution was added, and the mixture was reacted at room temperature for 20 minutes. After the reaction is complete, the suspended matter is filtered off and purified with IN-hydrochloric acid.
H-3 was prepared and 3,9-dicarboxyl-4'-
N-(β.

β,β-trichloroethoxycarbonyl)staurosporine was precipitated. This substance was purified by silica gel column chromatography (elution solvent: chloroform-methanol-acetic acid) (yield 60%).

HNMR (90Mllz, DMSO-d6δ)
:9.97 (s, ltl, ), 8.72 (s
, IH), 8.61 (s, 01), 8.
10 (d.

Itl, J=8.611z), 8.07 (d
, 111. J=8.6tlz), 8.02 (
d, 1!1, J=8.611z), 7.73(d,
Ill, J=8.611z), 7.12(br,s I
H), 5.15 (s, 211), 4.90 (s, 211)
), 4.15 (br, s, IH), 3°2~2.2
(m, 3H), 2.61 (s, 3H), 2.45 (s,
3! l) 1.96 (s311) I R (K B r ): 3400,2930,
1683,1585.1457cm-'MS m/z
: 728 (M'), 730 (M'+
2)732(M''+4) (iii) 3.9-dicarboxyl staurosporine 3.9-dicarboxyl-4'-N-(β, β5β
-trichloroethoxycarbonyl)staurosporine 3
6Hw (0.05mmo 1) to methyl cellosolve 1
After dissolving in 6ml, 4.6g of zinc powder and 2N-
2.5 ml of hydrochloric acid was added, and the mixture was stirred at room temperature for 30 minutes. After the reaction was completed, zinc was filtered off and fgIIX was removed under reduced pressure to obtain a syrupy liquid. This liquid was analyzed by HPLC (column: Y gate C-PACK S-3450O3, λ-294 nm
Solvent: C1l, C00NI1. aq-MeOH) and further desalting to obtain 8.2■ of 3,9-dicarboxyl staurosporine (yield: 3
1%).

H-NMR (400Mtlz, DMSO-db+
DzOδ): 9.96 (d, lfl, J = 1.6 Hz
), 8.52 (d, 18.J=1.6 fiz), 8.
08 (dd, lit, J=1.6 Hz, 8.7flz
), 8.07 (d, 111.J=8°911z), 8.
02 (dd, 01.J=1.6 Hz, 8.9 Hz),
7.70 (dIII, J=8.711z), 6.78
(d, 18.J=4.211z), 5.03(s.

2fl), 4.11(d, III, J=3.6 Hz
), 3.38 (s, 311), 3.29 (br, s, 1
8), 2.4-2.2 (m, 2K), 2.32 (s, 3
11), 1.36 (s, 3H) ”C-NMR (400MHz, DMSO-d6+[1
20δ) 171.838.168.407.167.9
26.142.199°138.765,132.79
2.130.895.128.480°127.682
．． 126.500, 125.760.123.614゜122.735.122.436.122.097, 1
21.887゜119.734.115.290.11
4.945.114.132゜108.418. 91
．． 381. 82.660. 80.126°57.2
58. 49.683. 45.442. 33.22
1°29.935. 29.174 I R (K B r ): 3420,1676.
1592.1542, 1473.1376, 1294.
1225cm M5 m/z: 554 (M”) Example 3 3.9-di(methoxycarbonyl)staurosporine 3.9-dicarboxylstaurosporine 100mg
(0,18 mmo 1> to methyl cellosolve 50 rn
2 ml of IOM-diazomethane in dichloromethane 18 was added, and the mixture was reacted at room temperature for 1 H.

After adding acetic acid to the reaction-completed solution to decompose excess diazomethane, the solvent was removed under reduced pressure to obtain 90μ of 3,9-di(methoxycarbonyl)staurosporine (8
6%). This compound can be purified by silica gel column chromatography (chloroporum-methanol) or recrystallization from chloroform.

'HNMR (400MHz, DMSO-dh, δ
): 9.97 (s, 1ll), 8.64 (s, 1
11), 8.51 (s, LH), 8.15-8.00 (
m, 3H), 7.72 (d, IH, J=8.5H
z), 6.78 (s, 1ft), 5.03 (s,
211), 4.10 (s, IH), 3.92 (s, 6H
), 3.38 (s.

3tl), 3.27(s, III), 2.4-2.2(
m, 2H), 2.32 (s, 311) 1.31 (s, 3
H) ” CNMR (400MHz, DMSO-dh
δ) 171.729.167.227.166.75
7.142.375°138.851.132.846
．． 130,919.128.226127.727.
126.222. 125.460. 123.614
122.562.122.084.121.120.1
20.722119.799.115.531.114
．． 927.114.100°108.635. 91.
348. 82.670. 80.10057.169
．． 52.105. 51.971. 49.547°45.460. 33.196. 29.954 29
．． 104I R (K B r ): 3450. 1
704. 1690. 1645. 1588°146
0, 1438.1356.132B, 12B2.125
8.1150.772.744cm-'

Claims (2)

[Claims] (1) General formula (I) ▲ Numerical formulas, chemical formulas, tables, etc. , and R and R^1 may be the same or different. However, R and R^1 cannot both be hydrogen.) A staurosporine derivative and a salt thereof. (2) General formula (II) ▲There are mathematical formulas, chemical formulas, tables, etc.▼(II) (In the formula, R^2 and R^3 represent hydrogen, formyl group, and carboxyl group, and R^4 represents hydrogen, R A staurosporine derivative represented by ^5CO group, and R^5 represents an alkyl group having 1 to 3 carbon atoms.