PROTEIN PHOSPHATASE INHIBITOR

Abstract: Novel protein phosphatase inhibitor and protein phosphatase inhibiting reagent, each of which comprises at least one physiologically active substance selected from the group consisting of NK374186 represented by chemical formula (I) and derivatives thereof or a pharmacologically acceptable salt thereof as an active ingredient.

Description: DESCRIPTION PROTEIN PHOSPHATASE INHIBITOR Technical Field This invention relates to a protein phosphatase inhibitor. Since dephosphorylation of proteins is a fundamental mechanism for regulating biological functions, an inhibitor thereof is useful as a drug for treating diseases caused by abnormalities in dephosphorylation or a reagent for inhibiting dephosphorylation of proteins. Background Art It is known that phosphorylation and dephosphorylation of proteins .in vivo are one of fundamental mechanisms for regulating biological functions. There have been reported a number of drugs capable of inhibiting phosphorylation of proteins, and it is also known that some of them show carcinostatic effects. In contrast, there have been reported few examples of protein phosphatase inhibitors [e.g., okadaic acid, Proc. Natl. Acad. Sci., USA, 85, 1768 - 1771 (1988)] which also play an important role in the proliferation and differentiation of cells. The above-mentioned okadaic acid would not be clinically used because of its high cytotoxicity and the like. A protein phosphatase inhibitor is useful as a drug for treating diseases in which a protein phosphatase is involved. It is also useful as a reagent to be used in, for example, clarifying the mechanisms of cell proliferation and differentiation and studying the function of a protein phosphatase. Accordingly, it is an object of the present invention to provide a novel protein phosphatase inhibitor. It is another object of the present invention to provide a protein phosphatase inhibiting reagent. Disclosure of Invention The present invention relates to a protein phosphatase inhibitor which comprises at least one physiologically active substance selected from the group of consisting of NK374186A, NK374186B, NK374186B3, NK374186C3, NK374186A3, NK374186D3, NK374186E3, NK374186H, NK374186I, NK374186P and NK374186PP represented by the following chemical formulae or a pharmacologically acceptable salt thereof as an active ingredient, together with a pharmaceutically acceptable carrier: NK374186A: NK374186B: NK374186B3: NK374186C3: NK374186A3: NK374186D3 : NK374186E3: NK374186H: NK374186I: NK374186P : NK374186PP ; The present invention also relates to a protein phosphatase inhibiting reagent which comprises at least one physiologically active substance as described above or a pharmacologically acceptable salt thereof as an active ingredient. The present invention also relates to a method for inhibiting a protein phosphatase which comprises administering to a mammal at least one physiologically active substance as described above or a pharma¬ cologically acceptable salt thereof. The present invention also relates to use of at least one physiologically active substance as described above or a pharmacologically acceptable salt thereof for the preparation of a protein phosphatase inhibitor. The present invention also relates to use of at least one physiologically active substance as described above or a pharmacologically acceptable salt thereof for the preparation of a protein phosphatase inhibiting reagent. Brief Description of Drawings Fig. 1 shows the ultraviolet absorption spectrum of NK374186A3. Fig. 2 shows the proton nuclear magnetic resonance spectrum of NK374186A3 as measured in dimethyl sulfoxide-dβ. Fig. 3 shows the carbon-13 nuclear magnetic resonance spectrum of NK374186A3 as measured in dimethyl sulfoxide-dβ- Fig. 4 shows the ultraviolet absorption spectrum of NK374186D3. Fig. 5 shows the proton nuclear magnetic resonance spectrum of NK374186D3 as measured in dimethyl sulfoxide-dβ. Fig. 6 shows the carbon-13 nuclear magnetic resonance spectrum of NK374186D3 as measured in dimethyl sulfoxide-dβ * Fig. 7 shows the ultraviolet absorption spectrum of NK374186E3. Fig. 8 shows the proton nuclear magnetic resonance spectrum of NK374186E3 as measured in dimethyl sulfoxide-dβ. Fig. 9 shows the carbon-13 nuclear magnetic resonance spectrum of NK374186E3 as measured in dimethyl sulfoxide-dβ* Best Mode for Carrying Out the Invention The physiologically active substance of the present invention is at least one physiologically active substance selected from the group consisting of NK374186A, NK374186B, NK374286B3, NK374186C3, NK374186A3, NK374186D3, NK374186E3, NK374186H, NK374186I, NK374186P and NK374186PP (hereinafter sometimes referred to as NK374186s) represented by the following chemical formulae or a pharmacologically acceptable salt thereof. The NK374186S according to the present invention are represented by the following chemical formulae. NK374186A: NK374186B : NK374186B3 NK374186C3: NK374186A3 NK374186D3; NK374186E3: NK374186H: NK374186I: NK374186P : NK374186PP: The physiologically active substances NK374186A, NK374186B, NK374186B3 and NK374186C3 can be produced in accordance with the method as described in Japanese Patent KOKAI (Laid-Open) No. 194571/1993; ϋSP. No. 5,306,496 and EP-A-0525361. More specifically, these substances can be obtained by culturing a NK374186 strain, which belongs to the genus Penicillium and is capable of producing the above-mentioned physiologically active substances NK374186A, NK374186B, NK374186B3 and NK374186C3, and recovering the physiologically active substances thus accumulated in the culture medium. The NK374186 strain was deposited in Fermentation Research Institute Agency of Industrial Science and Technology (Ibaraki-ken, Japan) on June 1, 1991 and given Accession Number FERM P-12285, and then transferred into an international deposition under the Budapest Treaty on May 25, 1992 and given Accession Number FERM BP-3870. Similarly, the physiologically active substances NK374186A3, NK374186D3 and NK374186 E3 can be obtained by culturing the above-mentioned strain and recovering these physiologically active substances thus accumulated in the culture medium. On the other hand, the physiologically active substances NK374186H, NK374186I, NK374186P and NK374186PP can be produced by the chemical conversion method by the use of NK374186B as a starting material, which is des¬ cribed in Japanese Patent KOKAI (Laid-Open) No. 194571/ 1993; USP. No. 5,306,496 and EP-A-0525326. These physio- logically active NK374186s may be in the form of pharma¬ cologically acceptable salts thereof. Examples of such salts include sodium, potassium, calcium and ammonium salts. The protein phosphatase inhibiting activities of the physiologically active substances NK374186S will be described below. The protein phosphatase inhibiting activities of the physiologically active substances NK374186S have been examined by using VHR protein phosphatase (Toshio Ishibashi et al., Proc. Natl. Acad. Sci. USA 89, 12170- 12174, 1992) and protein tyrosine phosphatase CD45 (Tonks, N.K. et al.. Biochemistry, 27, 8695-8701, 1988) each as a protein phosphatase deried from human cells. As a result, each of NK374186s shows an inhibitory activity on those protein phosphatases. The toxicities of the physiologically active substances NK374186s will be described below. Each of the physiologically active substances NK374186s was intraperitoneally administered to male CDF- mice aged 8 weeks in a dose of 100 mg/kg once a day continuously for 10 days. Each of NK374186S was dissolved in a mixture comprising 3.5% of DMSO, 6.5% of Tween 80 and 90% of a physiological salt solution, and then vialed. After monitoring for 10 days, none of the NK374186s caused any case of death. The physiologically active substances NK374186s significantly inhibit protein phosphatase playing impor¬ tant roles in the proliferation and differentiation of cells, which makes these substances useful as inhibitors of these enzymes and drugs for treating diseases involved in protein phosphatase reactions. Moreover, they are useful as reagents to be used in, for example, clarifying the mechanisms of cell proliferation and differentiation and studying the function of protein phosphatase. When the physiologically active substances NK374186S are to be used as drugs, they may be formulated into medicinal preparations and administered in accordance with methods conventionally used in the art. Namely, examples of the administration method include parenteral injection, oral administration and rectal administration. Examples of the dosage form usable therefor include injec¬ tions, powders, tablets and suppositories. To formulate these preparations, use can be made of various pharmaceu¬ tically usable carriers (for example, fillers and other auxiliary ingredients such as stabilizers, antiseptics, soothing agents and emulsifiers) if necessary, so long as they have no harmful influence on the NK374186s. In the formulation, the content of the physio¬ logically active NK374186s may vary over a wide range depending on, for example, the dosage form. In general, such a preparation comprises from 0.01 to 100% by weight, preferably from 0.1 to 70% by weight, of the NK374186S together with carriers (fillers, etc.) conventionally used in drugs as the balance. The NK374186s may be administered to an adult in a dose of from about 0.01 to 800 mg per day, though the dose varies depending on, for example, the conditions of the patient. When chronic administration is needed, it is preferable to decrease the daily dose. When the physiologically active substances NK374186S are used as a reagent, for example, they are dissolved in an organic solvent such as dimethyl sulfoxide or a surfactant, diluted with various buffer solutions, for example, Tris buffer solution, and then used in enzyme reactions and the like. To further illustrate the present invention in greater detail. Examples will be given below. However, it is intended that the present invention be not restricted thereto but rather be construed broadly within its spirit and scope. Example 1 Inhibition test on protein phosphatase VHR derived from human cell The inhibitory activities of the NK374186s were determined in accordance with the following method. Namely, a protein phosphatase derived from human cells was prepared from Escherichia coli having a VHR protein phosphatase gene introduced thereinto. To this protein phosphatase were added the NK374186S diluted serially. As a substrate, p-nitrophenyl phosphate (mfd. by Sigma) was employed and reacted with the enzyme at 37 β C for 30 minutes. After the completion of the reaction, 1 N sodium hydroxide was added and the absorbance of the p-nitrophenol thus formed was measured with an EIA reader (Model 2550, mfd. by Bio-Rad). The data thus obtained were employed as an index for the judgement of the activity of the NK374186S. Then the 50% inhibitory concentrations were calculated and listed in Table 1 described below. As Table 1 shows, the NK374186S exhibit inhibitory activities on the protein phosphatase VHR derived from human cells. Example 2 inhibition test on protein tyrosine phosphatase CD45 By using a protein tyrosine phosphatase CD45 prepared from the cell membrane of cultured cells Ball-1, the inhibitory activities of the NK374186s were determined by the same method as the one described in the above Example 1. Then the 50% inhibitory concentrations were calculated and listed in Table 1 described below. As Table 1 shows, the NK374186P and NK374186PP exhibit inhibitory activities in particular on the protein tyrosine phosphatase CD45. Table 1 Inhibitory activities of NK374186s on protein phosphatase IC50 (μg/mL) NK374186 VHR CD45 A 2.0 >600 A3 2.7 >600 B 13.0 >1000 B3 9.0 >1000 C3 1.0 >1000 D3 1.2 >1000 E3 8.0 >1000 H 17.0 >1000 I 7.5 >1000 P 3.0 450 PP 15.0 420 Next, methods for preparing the NK374186s to be used in the present invention will be described below. Referential Example 1 Method for producing NK374186A3, NK374186D3 and NK374186E3 Fermentation One hundred ml of a seed culture medium having the composition as specified in the following Table 2 was pipetted into a 500 ml Erlenmeyer flask and sterilized in an autoclave at 120°C for 20 minutes. Subsequently, it was inoculated with one platinum loopful of the NK374186 strain (FERM BP-3870), which was then incubated therein at 27°C under rotating at 200 rpm for 2 days. The culture medium thus obtained was referred to as the primary seed culture. Table 2 Seed culture composition (wt. %) glucose 2.0 sucrose 1.0 lactose 1.0 glycerol 0.2 soybean meal 2.0 polypeptone 0.5 sodium nitrate 0.2 magnesium sulfate 0.1 silicone 0.05 Toho No. 1 0.03 tap water The secondary seed culture was prepared in the following manner. Into a 30 1 jar fermenter was introduced 20 1 of a medium having the same composition as the one employed for the primary seed culture followed by sterilization. Then 200 ml of the primary seed culture obtained above was aseptically transplanted thereinto and incubated therein at 25°C, while aerating at a rate of 20 1/min and agitating at 250 rpm for 3 days. The culture thus obtained was referred to as the secondary seed culture. The main incubation was carried out in the following manner. One hundred and fifty little of a production medium, which had the same composition as the one of the seed culture medium but contained 1.0% of glycerol, was fed into a 200 1 tank and sterilized. Then 2 1 of the secondary seed culture obtained by the above method was aseptically transplanted thereinto and incubated at 25°C, while aerating at a rate of 150 1/min and agitating at 250 rpm for 4 days. Culture media obtained from 4 tanks (200 1) were filtered by using a filter press to thereby separate the cells from the filtrate. Purification To the cells thus harvested was added 200 1 of methanol. After extracting under stirring for 3 hours, the cells were separated from the extract by filtration under reduced pressure. To the extract thus obtained was added the same amount of water. The resulting solution was poured onto a Diaion HP-20 column (20 1). After washing with 50% of methanol and eluting with 20 1 of 60% acetone, a fraction 1 was obtained. After subsequently eluting with 30 1 of 80% acetone, a fraction 2 was obtained. The fraction 1 was evaporated to dryness and then dissolved in 0.2 1 of water. The resulting solution was poured onto a Diaion HP-20ss column (2 1) and developed by linear gradient elution with 50 - 70% acetone to thereby give a fraction IA and another fraction IB in this order. These fractions IA and IB were each poured onto a Sephadex LH-20 column which had been equilibrated with chloroform/methanol (1 : 1). Thus, 800 mg of NK374186A3 and 130 mg of NK374186D3 were obtained, respectively, from the fraction IA and the fraction IB. Meanwhile, the fraction 2 was evaporated to dryness, poured onto a silica gel column (5 1) and washed with 5 1 of chloroform. Then, it was developed with chloroform/methanol (50 : 1) to thereby give a fraction 2A. This fraction 2A was then poured suc¬ cessively onto a silica gel column and a Sephadex LH-20 column each developed with hexane/acetone (3 : 1). Thus, 500 mg of NK374186E3 was obtained. The physico- chemical properties of these components were as follows. NK374186A3: MW = 735 (FAB-negative spectrum). Fig. 1 shows its ultraviolet absorption spectrum. Fig. 2 shows its proton nuclear magnetic resonance spectrum as measured in dimethyl sulfoxide-dβ, and Fig. 3 shows its carbon-13 nuclear magnetic resonance spectrum as measured in dimethyl sulfoxide-dβ. NK374186D3: MW = 675 (FAB-negative spectrum). Fig. 4 shows its ultraviolet absorption spectrum. Fig. 5 shows its proton nuclear magnetic 5 resonance spectrum as measured in dimethyl sulfoxide-dβ, and Fig. 6 shows its carbon-13 nuclear magnetic resonance spectrum as measured in dimethyl sulfoxide-dβ. NK374186E3: MW = 595 (FAB-negative spectrum). ° Fig. 7 shows its ultraviolet absorption spectrum. Fig. 8 shows its proton nuclear magnetic resonance spectrum as measured in dimethyl sulfoxide, and Fig. 9 shows its carbon-13 nuclear magnetic resonance spectrum as measured in dimethyl sulfoxide-dβ. 5 Referential Example 2 Preparation of NK374186H NK374186B (42.6 mg, 0.065 mmol) was dissolved in pyridine (6.5 ml) and acetic anhydride (30.7 μl, 0.325 mmol) was added thereto. The obtained mixture was stirred at room temperature over day and night. Then 0 the reaction mixture was concentrated under reduced pressure. The residue thus obtained was purified by silica gel column chromatography (silica gel 5 g, eluted with 2% methanol/chloroform) to thereby give NK374186H (36.0 g, 79%). Table 3 shows the 1H-NMR 5 and FAB-MS data of NK374186H. Table 3 1H-NMR (200MHz, DMSO-d 6 )δ: 8.31(1H, d), 8.18(1H, d), 7.95(1H, d), 5.42(1H, d), 5.22(1H, m) , 4.90(1H, m) , 4.67(1H, m), 4.54(2H, m) , 3.98(2H, m) , 3.62(1H, m), 2.22(1H, m) , 2.07(1H, m) , 2.00(3H, s), 1.95(3H, s), 1.71(1H, m), 1.50(2H, m), 1.23(22H, m), 1.12(6H, m) , 0.89(9H, m), 0.76(3H, d) FAB-MS m/z: 698[(M+H) + ] Referential Example 3 Preparation of starting compound (bl) for preparing NK374186I Compound (bl) : NK374186B (52.7 mg, 0.080 mmol) and 4- dimethylaminopyridine (10.1 mg, 0.083 mmol) were dissolved in methylene chloride (0.7 ml). In an argon atmosphere at -40°C, triethylamine (15.0 μl, 0.108 mmol) and methanesulfonyl chloride (7.4 μl, 0.0096 mmol) were added thereto. The resulting mixture was stirred under ice-cooling for 1.5 hours, and then triethylamine (15.0 μl, 0.108 mmol) and methanesulfonyl chloride (7.4 μl, 0.096 mmol) were further added thereto at -40 β C. The resulting mixture was stirred at 5°C overnight. Then a saturated aqueous solution of sodium hydrogencarbonate was added to the reaction mixture followed by extraction with methylene chloride. The organic layer thus obtained was washed with a saturated aqueous solution of sodium chloride and dried over anhydrous magnesium sulfate. Then the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (silica gel 8 g, eluted with 0 - 2% methanol/chloroform) to thereby give the above-mentioned compound (bl) (45.6 mg, 78%). The result of FAB-MS was as follows; m/z : 734 [(M+H) + ]. Referential Example 4 Production of NK374186I The above-mentioned compound (bl) (45.0 mg, 0.061 mmol) was dissolved in tetrahydrofuran (0.7 ml). Then l,8-diazabicyclo[5.4.0]undecene (10.1 μl, 0.067 mmol) was added thereto at room temperature and stirred. After 1.5 hours have passed, 1,8-diazabicyclo[5.4.0]- undecene (5.1 μl, 0.034 mmol) was further added thereto and the resulting mixture was stirred for 4 hours. After adding ethyl acetate, the reaction mixture was washed successively with a 1 N aqueous solution of hydrochloric acid, water and a saturated aqueous solution of sodium chloride and dried over anhydrous magnesium sulfate. Then the solvent was distilled off under reduced pressure and the obtained residue was purified by silica gel column chromatography (silica gel 3 g, eluted with 25 - 35% ethyl acetate/ hexane) to thereby give NK374186I (21.4 mg, 61%). Table 4 shows the 1H-NMR and FAB-MS data of NK374186I. Table 4 1H-NMR (200MHz, CDC1 3 )6: 8.15(1H, brs), 7.77(1H, brs), 6.97(1H, q), 6.63(1H, s), 6.35(1H, m), 5.98(1H, s), 4.91(1H, brs), 4.28(1H, m), 3.83(1H, m) , 3.42(1H, dd), 2.28(2H, m), 1.87(1H, m) , 1.80(3H, d), 1.40(2H, m), 1.24(22H, m) , 0.96(6H, m), 0.87(6H, m) FAB-MS m/z: 578[(M+H) + ] Referential Example 5 Preparation of starting compound (cl) for preparing NK374186P Compound (cl) : NK374186B (61.2 mg, 0.093 mmol) and 1H- tetrazole (32.6 mg, 0.465 mmol) were dissolved in methylene chloride (2 ml). After adding N,N-dimethyl- l,5-dihydro-2,4,3-benzodioxaphosphepin-3-amine (24.1 μl, 0.112 mmol) in an argon atmosphere, the resulting mixture was stirred at room temperature. Thirty minutes after, N,N-dimethy1-1,5- dihydro-2,4,3-benzodioxaphosphepin-3-amine (12.0 μl, 0.056 mmol) was further added and the resulting mixture was stirred for 20 minutes. After adding water (50 μl) and stirring for 10 minutes, the mixture was cooled to -40 β C and 3-chloroperbenzoic acid (140 mg, 0.811 mmol) was added thereto. After returning to room temperature, the reaction mixture was stirred for additional 10 minutes. Then a 10% aqueous solution of sodium sulfite (10 ml) was added to the reaction mixture. After stirring, it was extracted with ethyl acetate. The organic layer thus obtained was washed successively with a saturated aqueous solution of sodium hydrogencarbonate and a saturated aqueous solution of sodium chloride and dried over anhydrous sodium sulfate. Then the solvent was distilled off under reduced pressure and the obtained residue was purified by silica gel column chromatography (silica gel 15 g, eluted with 1% methanol/chloroform) to thereby give the compound (cl) (39.4 mg, 51%). The result of FAB-MS was as follows; m/z: 838 [(M+H) + ], 860 [(M+Na) + ]. Referential Example 6 Preparation of NK374186P The above-mentioned compound (cl) (29.1 mg, 0.035 mmol) was dissolved in an 80% aqueous solution of ethanol (5 ml) and 10% palladium-carbon (moisture content 50%, 10 mg) was added thereto. Then the mixture was stirred for 40 minutes while bubbling hydrogen gas thereinto. The reaction mixture was filtered through celite to thereby eliminate the palladium-carbon. To the filtrate thus obtained was added concentrated aqueous ammonia (0.1 ml) and the resulting mixture was concentrated under reduced pressure. Then the residue was dissolved in water (1 ml) and freeze-dried to thereby give NK374186P in the form of the ammonium salt thereof (25.9 mg, 96%). Table 5 shows the iH-NMR and FAB-MS data of the NK374186P. Table 5 IH-NMR (200MHz, DMSO-d6)6: 8.18(1H, d), 7.94(1H, d), 7.88(1H, d), 4.90(2H, m), 4.60(2H, m) , 4.35(2H, m) , 3.99(2H, m), 3.59(1H, m) , 2.12(2H, m), 2.00(3H, s), 1.60(3H, m) , 1.24(22H, m), 1.12(6H, m), 0.87(9H, m), 0.72(1H, d) FAB-MS m/z: 758[ (M+Na) + ] 780[(M+2Na) + ] Referential Example 7 Preparation of starting compound (c3) for preparing NK374186PP Compound (c3) : NK374186B (65.5 mg, 0.100 mmol) and 1H- tetrazole (50.0 mg, 0.714 mmol) were dissolved in methylene chloride (2 ml). After adding N,N-dimethyl- 1,5-dihydro-2,4,3-benzodioxaphosphepin-3-amine (0.104 ml, 0.482 mmol) in an argon atmosphere, the resulting mixture was stirred at room temperature for 15 minutes. After adding water (0.05 ml), the mixture was stirred for 15 minutes and cooled to -40 β C. Then 3-chloro- perbenzoic acid (148 mg, 0.858 mmol) was added thereto. After returning to room temperature, the reaction mixture was stirred for additional 15 minutes. Then a 10% aqueous solution of sodium sulfite (10 ml) was added to the reaction mixture. After stirring, it was extracted with ethyl acetate. The organic layer thus obtained was washed successively with a saturated aqueous solution of sodium hydrogencarbonate and a saturated aqueous solution of sodium chloride and dried over anhydrous sodium sulfate. Then the solvent was distilled off under reduced pressure and the obtained residue was purified by silica gel column chromatography (silica gel 15 g, eluted with 2% methanol/chloroform) to thereby give the above-mentioned compound (c3) (93.8 mg, 93%). The result of FAB-MS was as follows; m/z: 1020 [(M+H) + ], 1042 [(M+Na) + ]. Referential Example 8 Preparation of NK374186PP The above-mentioned compound (c3) (80.0 mg, 0.079 mmol) was dissolved in a 75% aqueous solution of ethanol (20 ml) and 10% palladium-carbon (moisture content 50%, 40 mg) was added thereto. Then the mixture was stirred for 1 hour while bubbling hydrogen gas thereinto. The reaction mixture was filtered through celite to thereby eliminate the palladium- carbon. To the filtrate thus obtained was added concentrated aqueous ammonia (0.6 ml) and the resulting mixture was concentrated under reduced pressure. Then the residue was dissolved in water (1 ml) and freeze-dried to thereby give NK374186PP in the form of the ammonium salt thereof (65.1 mg, 93%). Table 6 shows the 1H-NMR and FAB-MS data of the NK374186PP. Table 6 1H-NMR (200MHz, DMSO-d 6 )δ: 9.64(1H, d). 8.65(1H, d). 7.82(1H, d). 4.70-4.00(8H , m), 2.23(3H, m). 1.93(3H, s). 1.80(1H, m). 1.50(2H, m). 1.32(6H, m). 1.24(22H, m). 1.01(3H f t). 0.83(9H, m) FAB-MS m/z: 838[(M+Na) + ] 860[(M+2Na) + ] 882[(m+3Na) + ] Industrial Applicability According to the present invention which has been described in detail hereinabove, NK374186S or pharmacologically acceptable salts thereof are useful as protein phosphatase inhibitors, protein phosphatase inhibiting reagents, and the like.

Claims: CLAIMS 1. A protein phosphatase inhibitor which comprises at least one physiologically active substance selected from the group consisting of NK374186A, NK374186B, NK374186B3, NK374186C3, NK374186A3, NK374186D3, NK374186E3, NK374186H, NK374186I, NK374186P and NK374186PP represented by the following chemical formulae or a pharmacologically acceptable salt thereof as an active ingredient, together with a pharmaceutical¬ ly acceptable carrier: NK374186A: NK374186B: NK374186B3 NK374186C3: NK374186A3: NK374186D3: NK374186E3: NK374186H: NK374186I: NK374186P; NK374186PP; 2. A protein phosphatase inhibiting reagent which comprises at least one physiologically active substance according to Claim 1 or a pharmacologically acceptable salt thereof as an active ingredient. 3. A method for inhibiting a protein phosphatase which comprises administering to a mammal at least one physiologically active substance according to Claim 1 or a pharmacologically acceptable salt thereof. 4. Use of at least one physiologically active substance according to Claim 1 or a pharmacologically acceptable salt thereof for the preparation of a protein phosphatase inhibitor. 5. Use of at least one physiologically active substance according to Claim 1 or a pharmacologically acceptable salt thereof for the preparation of a protein phosphatase inhibiting reagent.

Inventors: MORINO TOMIO, OSADA HIROYUKI, MORINO, TOMIO, OSADA, HIROYUKI
Applicants: MORINO TOMIO, NIPPON KAYAKU KK, OSADA HIROYUKI, RIKAGAKU KENKYUSHO, MORINO, TOMIO, NIPPON KAYAKU KABUSHIKI KAISHA, OSADA, HIROYUKI, THE INSTITUTE OF PHYSICAL AND CHEMICAL RESEARCH (R
Application year: 1996
Main classification: C07K5
IPC codes: A61K38, A61P43, C07K5, C07K14, C12P21, C12R1