WO1992012968A1 - Pharmaceutical composition for thrombus - Google Patents

Abstract

1-Deoxymannojirimycine, a compound of formula (I), which is excellent in an α2-plasmin inhibitory activity and serves as a thrombolytic agent which is efficacious in treating a wide variety of diseases caused by thrombus, such as myocardial, cerebral and pulmonary infarctions.

Description

 Harm

 Thromboembolic drug composition

 Technical field

 The present invention relates to a pharmaceutical composition for thrombus.

 Natsume technique

 When a blood vessel is physically damaged and bleeds, the body quickly aggregates blood at the site of impairment of the blood vessel at the site of damage to the blood vessel to preserve life, and rapidly solidifies the blood through precipitation and polymerization of fibrin to form a thrombus. . However, since this hemostatic plug, on the other hand, impedes blood flow and causes ischemia and necrosis of the tissue at its peripheral site, the living body has a mechanism to remove excess thrombus and unnecessary thrombus. . The dissolution of excess or unnecessary thrombus is caused by the activation of brassinogen by a brassminogen activator (hereinafter referred to as “PAJ”), and the resulting brassin is converted into fibrin. Is caused by decomposition (fibrinolysis).

 On the other hand, if the thrombus is dissolved before the vascular tissue is completely restored, the living body loses blood and enters a dangerous state for maintaining life. For this reason, living organisms also have a mechanism to control excess fibrinolysis. 2-Brassin inhibitor-1 (hereinafter referred to as “α2-α”) plays a central role in this mechanism. << 2-ΡΙ inhibits the action of the above-mentioned plasmin in blood at the time of dissolution, prevents dissolution of the hemostatic plug, and stabilizes the thrombus by being taken into the hemostatic plug. In addition, α 2-ΡΙ increases after surgery as a kind of acute-phase protein, and is one of the causes of post-operative thrombus.

Myocardial infarction, cerebral infarction, pulmonary infarction, etc. Blood clots are thought to occur because they block blood flow and damage peripheral tissues. This is presumably because the balance between the fibrinolytic system and the coagulation system, which the living body originally has, has been lost. At present, thrombolytic therapy that administers PAs such as peroxidase (hereinafter referred to as ruKj), streptokinase, and krillogen plasminogen activator (referred to as "t-PAj" below) is used to cure these conditions. Is being done.

 However, as described above, α2-ΡΙ present in blood and thrombus inhibits plasmin produced by ΡΑ administration, so there is a natural limit to dissolving thrombus by 投 与 administration, and systemic There is a significant risk associated with side effects such as bleeding from sexual fibrinolysis.

 Therefore, if the activity of << 2- 血 in the blood can be reduced and the balance of the fibrinolytic system can be tilted more in the direction of fibrinolysis, it will help prevent postoperative thrombosis and thrombosis, as well as prevent myocardial infarction and It will be used to cure conditions such as pulmonary infarction caused by blood clots.

 It is known that when a monoclonal π-nal antibody that inhibits 2-ΡΙ activity is added to a thrombus formed in a test tube, the thrombus is rapidly dissolved (Sakata et al., Blood, 74, 2692- 2697 (1989)).

 On the other hand, the compound of the present invention, which is a known compound, and its derivative are known as mannosidase inhibitors. However, no attempt has been made to further study and identify the specific pharmacological action of the compound of the present invention and to link it to commercialization as a pharmaceutical. (Bl bein, A.D .; PASBB J, Vol. 5, 3055-3063 (1991), JP-A-61-200967)

First, the present inventors focused on the action of α2-ΡΙ, and found that N-substituted moranolin has an action of reducing the activity of α2-ΡΙ. Apply for a patent showing that it has a promoting effect (Japanese Unexamined Patent Publication No. 1-250350).

 The present inventors have found a drug having a 2-PI activity lowering action that is stronger than the previously disclosed N- 厣 换 moranoline, and has been found to be a therapeutic agent for diseases such as myocardial infarction and pulmonary infarction that widely hide the cause of thrombus. The study was conducted for the purpose of providing a thrombolysis accelerator effective for the treatment.

 Disclosure of the invention

 In order to achieve the above object, the present inventors have screened a large number of compound groups and, fortunately, obtained 1-doxymanno jilimycin (^ below) represented by the following general formula [I]. MI C-1 J) and pharmacologically acceptable acid addition salts thereof have a stronger effect of lowering 2-PI activity than N-substituted moranolin. Was confirmed to be effective, and the present invention was finally completed.

[I]

Here, the acid-added clay includes inorganic acid salts such as hydrochloride, nitrate, sulfate, hydrobromide, and uranate, calcium carbonate, succinate, fumarate, and maleate. Organic acid salts such as acid salts, lingate salts, lactate salts, tartrate salts, and methansulfonate salts.

 It has been found that the compounds of the present invention have extremely low toxicity.

As shown in the test examples later, the compound of the present invention has a remarkable thrombolysis-promoting effect in vitro. The compound of the present invention is effective, for example, in diseases based on thrombus formation such as myocardial infarction, angina pectoris, cerebral infarction, pulmonary infarction, peripheral arteriovenous obstruction, and postoperative thrombus. However, for example, the drug effect can be exhibited more effectively by using it in combination with t-PA (human ssue-type plasminogen activator). The medicament of the present invention is extremely effective in preventing thrombosis recurrence.

 When the pharmaceutical composition of the present invention is administered, it can be applied in combination with or mixed with the drug to be administered simultaneously. The compound of the present invention is used alone or in a pharmaceutically acceptable nontoxic and inert carrier, for example, as a pharmaceutical composition containing 0.1% to 99.5%, preferably 0.5% to 90%. Administered to animals, including humans.

 As the carrier, a solid, half-faced or liquid diluent, filler, and one or more other prescription aids are used. Preferably, the pharmaceutical compositions are administered in dosage unit form. The pharmaceutical composition of the present invention can be administered by injection, intradermal administration, topical administration (eg, percutaneous administration), or rectally. Of course, it is administered in a type II suitable for these administration methods. For example, intravenous administration and intravenous administration, particularly intravenous administration, are preferred.

It is desirable to adjust the dose as an antithrombotic agent in consideration of the patient's condition such as age, body weight, etc., the administration route, the nature and extent of the disease, and the like. The amount of the active ingredient is generally in the range of 10 mg / day to: LO g / day Z, preferably 100 mg to 3 g / day. In some cases, less may be sufficient, and conversely, this may require a dose on JSi. It is also desirable to administer the drug in 2 to 3 divided doses a day. Peripheral administration is in the form of solid or liquid dosage units, for example, powders, powders, tablets, dragees, capsules, capsules, granules, fragrances, liquids, syringes, drops, sublingual tablets, etc. Can be carried out depending on the dosage form.

 Powders are prepared by shaping the active substance to a suitable consistency. Powders are prepared by commingling the active substance with a rapid fineness and then mixing with the similarly finely divided pharmaceutical carrier, for example, edible carbohydrates such as flour, mannitol and the like. If necessary, flavoring agents, preservatives, dispersing agents, coloring agents, flavors and the like may be mixed.

 The force busel is prepared by filling the powdered powder, powder or tablet granulated as described above into granules, as described in the section on tablets, into a capsule of capsule such as gelatin capsule. Manufactured. Lubricants and glidants, such as colloidal silica, talc, magnesium stearate, calcium stearate, and solid polyethylene glycol are mixed with the powdered material. However, the filling operation can be performed later. Disintegrants ゃ Solubilizers such as carboxymethylcellulose, carboxymethylcellulose TD-sCalcium, low-deposition hydroxylose vircellulose-sucrose, quascarmellose sodium, carboxymethyl starch sodium Addition of um, carbonated sodium and sodium carbonate can improve the efficacy of the medicine when the capsule is taken.

In addition, the powder of this product can be suspended and dispersed in vegetable oil, polyethylene glycol, glycerin, and a surfactant, and wrapped in a gelatin sheet to form a soft capsule. Tablets are manufactured by adding a shaping agent to form a powder mixture, granulating or slugging, then adding a disintegrant or lubricant and tableting. The powder mixture should be Mix the powdered substance K with the diluent or pace described above, and if necessary, combine with a binder (eg, sodium propyloxymethylcellulose, methylcellulose, hydroxypropyl pillmethylcellulose, gelatin, polyester). Vinylpyrrolidone, polyvinyl alcohol, etc.), dissolution retarders (eg, paraffin), resorbents (eg, tetrasalt) and adsorbents (eg, pentonites, kaolin, dicalcium phosphate) Etc.) may be used together. The powder mixture is first moistened with a binder such as syrup, starch paste, gum arabic, a cellulose solution or a high molecular weight solution, stirred and mixed, and dried and pulverized into granules. it can. Instead of granulating the powder in this way, it is also possible to first apply the powder to a punch and then crush the resulting imperfect form of the slag into granules.

 The granules thus produced can be prevented from adhering to each other by adding stearic acid, stearate, talc, mineral oil and the like as a lubricant. The lubricated mixture is then tableted. The uncoated tablets produced in this way can be coated with Filmco Single Coating.

Alternatively, the drug may be directly tableted after mixing with a fluid inert carrier without performing the steps of granulation and slag formation as described above. A transparent or translucent protective coating consisting of a sealing coat of silica, a coating of sugar molecular material, and a polishing coating consisting of wax can also be used. Other oral dosage forms such as solutions, syrups and elixirs can also be made into dosage unit form so that a given quantity contains a fixed amount of the drug. Syrup is made by dissolving the compound in an appropriate aqueous solution. Elixir is prepared using a non-toxic alcoholic carrier. It is manufactured by Suspensions are formulated by dispersing the compound in a non-toxic carrier. Solubilizers and emulsifiers (eg, ethoxylated isosteryl alcohols, polyoxyethylene sorbitol esters), preservatives, flavor enhancers (eg, palmit oil, saccharin) Others can also be added as needed.

 If necessary, the dosage unit formulation for microdose administration can be microbubbled.该 Formulations can also be extended in duration or sustained release by coating or embedding in polymers, waxes, etc.

 Intrathecal administration can be carried out by using a liquid dosage unit form for subcutaneous, intramuscular or intravenous injection, for example, a solution / turbidity form. These include dissolving or dissolving a fixed amount of the compound in a non-toxic liquid carrier suitable for the purpose of injection, such as aqueous or oily media, and then sterilizing the suspension or solution. It is manufactured by Non-toxic salts or salt solutions may be added to make the injection solution isotonic. In addition, stabilizers, preservatives, emulsifiers and the like can be used in combination.

 Rectal administration involves administering the compound in a low water soluble or insoluble facet, such as polyethylene glycol, cocoa butter, semi-synthetic fats and oils (such as Witebsol®), esters (such as palmitin). It can be carried out by using a suppository or the like prepared by dissolving or suspending in acid mixture I) stildester) and a mixture thereof. In the preparation of the compound of the present invention, other drugs such as UK, streptokinase, t-PA and the like can be used in addition to the active ingredient of the present invention.

Hereinafter, the present invention will be described in more detail with reference to test examples for the << 2-Ρ activity lowering action and thrombolytic action of the compound of the present invention. Test Example 1 Blood 2-PI activity lowering effect in beagle dogs (administration in gestation)

 1—Deoxymanno zilimycin (^ lower, called “MI C-1J”) reduced the serum α2-ΡΙ activity in 4 male beagle dogs (7-month-old, body weight 9 ~: llkg) using a group of N —Methylmoranoline (hereinafter referred to as “M0R-14”).

 The experiment was performed as follows.

 First, dissolve in sterile physiological saline so that the S degree of MIC-1 is 3, 10, and 30 flJg / ml, and the contact degree of M0R-14 is 100 fflg / ml. The solution was sterilized for tt using a sterile filter (point size 0.2 x ra), and 1 nil per kg of body weight of a beagle dog was administered once a day through the forelimb vein at 9:00 pm 3 days K. Sterile physiological saline was similarly administered to the control group.

 Blood samples were collected on the first day of administration (Day 0), the next day (Day 1), 6 hours after each drug administration on the last day (Day 2), and 24 hours after the last dose (Day 2) (Day 3). Went after K (Day 4). The collected blood was mixed with 1 volume of 3.8S5 sodium citrate to 9 volumes of blood, and centrifuged at 3000 rpra X 15 min to separate the plasma.

 ατ 2-ΡΙ activity was measured using a measurement kit test kit APL · 2 kit manufactured by Daiichi Pure Chemicals Co., Ltd.

Table 1 shows the measurement results. The values in parentheses are the values when the plasmin inhibitory activity before administration of the subject was set to 100. Table l Male 2-PI¾¾ «Ϊpulse! ¾δ given)

 ": Θϋ ^ from Ofiifim (hri

 *: P <a05 (Dunnett) for the control n

 **: P <a01 relative to control (Dunnett) As is clear from Table 1, administration of MIC-1 resulted in a dose-dependent decrease in a2-PI activity of # 8.

 At 3rog / kg IH C-1 dose, a 2-PI activity decreased to 72% of the pre-dose 30 hours ¾ after the first dose (p <0.05), and 48 hours and 54 hours later Then, they dropped to 51% and 58%, respectively (P <0.01). A significant lowering effect was also observed at 24 hours K (Day 3) after the last administration of the drug, but it recovered slightly after 48 hours (Day 4).

 At 10 mg / kg and 30 mg / kg MIC-1 gun administration, 2-PI activity decreased to 70% and 60%, respectively, 24 hours later than the pre-dose value (p <0.05), After 30 hours, they dropped to 59% and 52%, respectively (p <0.01). In the 10 nig / kg group, the values decreased to 50% and 39% after 48 and 54 hours, respectively, and in the 30 mg / kg group, they decreased to 38% and 34%, respectively. This value was still significantly reduced 24 hours after the last administration of the drug (Day 3), and recovered slightly after 48 hours K (Day 4).

At 100 rog / kg JJ0R-14 swift gun administration, α2-PI activity is 30 hours after the first dose Shortly thereafter, it decreased to 84% of that before administration (P <0.05), and decreased to 68% after 54 hours (P <0.01). After the administration, this 值 recovered promptly.

 Based on the results above £ 1, MI C-1 significantly reduced QT 2-PI activity compared to M0R-14, and at a dose 30 times lower than M0R-U, the activity of 2-ΡΙ was sufficient. To lower.

 Test Example 2 Reduced 2-ΡΙ activity in blood in beagle dogs (oral L

The effect of reducing the 2-ΡΙ activity in the blood by g oral administration of MIC-1 was measured using a group of five male beagle dogs (10-month-old, weighing 9-ll kg) in comparison with K0R-14.

 The experiment was performed as follows.

 The capsules were filled with JJIC-l (30 mg / kg) and M0R-U (300 mg / kg) at the respective doses and administered orally once daily at 3 pm on March 3rd.

 Blood samples were collected on the first day of administration (Day 0), the next day (Day 1) and before the last administration (Day 2), before drug administration, after 6 hours of administration, and 24 hours after the last administration (Day 2) (Day 3). After 48 hours (Day 4) and after 120 hours (Day 7). The collected blood was mixed with 1 volume of sodium sodium citrate to 9 volumes of blood, and centrifuged at 3,000 rpm x 15 rain to obtain plasma.

 The QT 2-PI activity was measured using a test team APL kit (Daiichi Pure Chemicals Co., Ltd.).

Table 2 shows the measurement results. The value in 0 is 值 when the plasmin inhibitory activity before administration of the test subject is set to 100.

 ": Mm and ffls before (« separation

 w :: From ras ^ ΦββΜ <hr: im

 t: P <0.05 for control (Dunnett)

 **: Control o—P <0.01 (Dunnett)

As is clear from Table 2, administration of MIC-1 resulted in a dose-dependent decrease in α2-ΡΙ activity in S8.

 Twenty-four hours after administration of MIC-1 (30 aig / kg) and MOR-l "300 ojg / kg), reductions were found to be 70% (p <0.01) and 86% (P <0.01), respectively. , «IC-Κ M0R-14 was reduced to 36% (p <0.01) and 47% (p <0.01) before administration, respectively, at 6 o'clock bending (54hr.) After administration of 3 days of sunshine. At 24 hours (72 hr.) And 48 hours (96 hr.) After administration, a2-PU gradually recovered, and after 120 hours (168 hr.), Both recovered to 80% to 90% of those before administration.

 Based on the above results, both MIC-1 and M0R-14 significantly reduced plasma a2-PI activity when administered in vitro, and MIC-1 was effective at lower doses compared to M0R-U. .

The same test was conducted for the compound described in JP-A-61-200967 (for example, N-methyl-11-deoxynojirimycin). As a result, in all cases, 30 mg / kg did not show any << 2- 低下 activity lowering effect o

 Test Example 3 Thrombolytic action in in vitro n (in vitro)

 Using the plasma obtained in Test Example 1, a fibrin mass was prepared in vitro, and the solubility of the thrombus dissolving agent lii (when acted on) was measured. -ΡΙ The activity of which showed a marked decrease in activity was subjected to the test, ie, for MIC-1 (3, 10, 30 rag / kg), 72 hours after the initial administration, and for MOR-U (100 ng / kg). For plasma, plasma obtained 54 hours after the initial administration was used.

 The experiment was performed as below J¾.

First, to the separated blood sample 500 1 was added 10 il of bubrinogen (a85kBq / ml) and mixed, and the mixture was dispensed at 100 / il into test tubes. Then, 25 ill / ml tombin solution and 0.5% calcium carbonate solution were added in 5 jul each, and the mixture was easily cupped at 37 "C for 30 minutes to produce fibrin 9 mass. A solution of UI (0, 10 and 30 UI / ιη1) in the culinary solution was added to each test tube with 1 oil, and the mixture was incubated at 37 tons, and 3, 6, 9, 12 and Twenty-four hours later, 25 xl of the supernatant was collected in a RU tube, and the radioactivity of the ^12S I-fibrin degradation product that migrated into the supernatant was measured using a 7 ″ -counter.

 Figures 1 and 2 show the 30 U / ml UI (temporal change in fibrin clot dissolution when 1 ml of the solution was added (average planting of 4 cases per group)).

 As is clear from the figure, the fibrin clot lysis rate obtained from dog plasma treated with MIC-1 (3, 10, 30 mg / kg) and MQR-U (1 OOrog / kg) was higher than that of control. Significantly increased.

Tables 3 and 4 show the dissolution rates 12 hours after (0U, 10U, 30U) addition. Shown in

 The parentheses in parentheses are %% of the control group.

 Table 3 Evelyn ®a¾? Sii (½)

 Control Τϊ'-le MIC-IS ^ group

 鲰 瞵 rank N 3mgAg lOnsks 30oskg

 4 29 ± 2 28 ± 1 (97) 28 ± 1 (97) 27 ± 1 (93) 4 3 ± 1 45 + 7034) 5? ± 6 '* (168) 80 ± 6 ** (235) 4 42+ 5 6 ± 10 (152) 87 ± 8 ** (207) 101 ± 1 ** (240)

 1! +++ 1

 <r2-PIS¾0) 4 89 ± 6 50 ± 8 4 ± 10 33 ± 4

 *: Control ΛΗ = ¾ and P <05 (Ounnett)

 **: 01 for control n (Dunnett)

Table 4 Fibrin from ΜΟίΗ ^ δ ^ nu © i ^? ³ ? (¾) Control

 N QOOng / kg)

 4'32 ± 2 019)

4 46 ± 3 * (135)

4 60 ± 5 * (146>

 «2-PI¾¾«) 4 89 ± 6 76 ± 5

*: Contrast ti vs P 0.05 (Ounnett) In the HI C-1 administration group, the fibrin dissolution rate without addition was the same as that in the control group, but when 10 U or 30 U of UK was added, MIC-1 was dose-dependently dependent on the fibrin dissolution rate. Was enhanced. That is, the fibrin dissolution rate of the MIC-1 administration group of 3 rag / kg, lOrag / kg, and 30 mg / kg increased by 34, 68, and 135 X, respectively, with respect to the control group when 10 U of UK was added. When 30 U of UK was added, the fibrin dissolution rate of the MIC-1 administration groups of 3 nig / kg, 10 mg / kg and 30 nig / kg increased by 52, 107 and U0X, respectively, as compared with the control group.

 In the 100 rag / kg M0R-14 administration group, a slight increase in dissolution was observed in the control group without the addition of UK, and the fibrin dissolution rate was lower than that in the control group when ϋΚ was added at 10 U or 30 U. 35, increased (P <0.05).

 ^ From the above results, it is clear that in plasma whose activity has been reduced by administration of MIC-1 and M0R-14, << dissolution of fibrin clots by ϋΚ increases according to the degree of decrease in 2-ΡΙ activity, and this effect Was particularly marked in the MIC-1 administration group.

Test Example 4 Thrombolysis promoting effect in a canine pulmonary infarction model

 Test Example 1, 2, and 3 show that MIC-1 administered to dogs for 3 consecutive days (intravenously, percutaneously) decreases << 2-ΡΙ activity in plasma, and thrombus produced using the plasma Has been found to dissolve more rapidly in the presence of oral kinase than control. Therefore, an experiment using a canine pulmonary infarction model was conducted in order to examine the thrombus dissolution promoting effect of MIC-1 in vivo.

Male beagle dogs (8 to 20 liters) given MI C-KlOffig / iBl / kg) or saline ice (loi l / kg) (control group) once a day for 3 days. Anesthetized with pentobarbital (0.6 rol / kg) at the age of 8 to 14 kg), a 5P sheath-introducer (manufactured by Terumo) is placed in the facial vein distal to the bifurcation of the jugular and facial veins. did. 1 volume of 3.8% sodium citrate and 9 volumes of blood were rubbed, and 2 ml of this blood was added to ¹²⁵ 1-fibrinogen (2.6 MBeq / ml) 10ju 1, 25 <nM CaCl ₂ solution and 50〃1 of thrombin (lOOU / ml) was sequentially added, and the mixture was inhaled into a polyethylene tube having an inner diameter of 3 rom. The formed thrombus is extruded from the tube, cut into a length of about 3cro, and washed with saline.

(20rolx 3).

 After measuring the total radioactivity of the thrombus with an r-line force center (A5424, manufactured by Packard), the thrombus was injected into the body through a sheath introducer. It was confirmed using a G1! Tubular survey meter (TGS-113, manufactured by AROKA) that the thrombus was retained in the peripheral region of the lungs through the jugular vein, right atrium, and right ventricle.

 t-PA was administered at 0.4 ng / 0.4ral / kg immediately after infusion of thrombus through Sheath transducer.

 During the observation period, physiological saline was continuously injected at a rate of 0.5 ml / rain through a sheathintroducer. At 0, 15, and 30 minutes after the injection of the thrombus, 2 nil of blood was temporarily collected every 30 minutes until 420 minutes thereafter, and the radioactivity was measured with an r-line force monitor. The radioactivity in whole blood at each measurement point was calculated based on the blood volume as 1/13 of body weight, and (radioactivity in whole blood / total radioactivity) X 100 (hereinafter referred to as "X of dose j") was calculated. The measurement results are indicated by the following four indices.

 (1) The highest% of dose dose (hereinafter referred to as “peak height”)

(2) The time when it was observed (hereinafter referred to as “peak time”) (3) Total of dose up to 420 minutes (hereinafter referred to as “X of dose total 楦”)

(4) When reaching the total of 50 ^ in (3) (when reaching 50 %%, hereafter referred to as “T ₅₀ ”). Table 5 shows the results.

Table 5 Thrombolytic activity in canine pulmonary infarction model

 Beak beak% of dose Τδ o group N total time height (min)

 (Min) (min) (up to 420 min) η- & 4 233 ± 8 19 ± 2 206 ± 20 223 ± 15 IC-1 4 128 ± 43 22 ± 2 252 ± 22 175 ± 14

(10rag / kg) (55) (116) (122) (78) t-PA 4 68 ± 38 ^b 26 ± 3 273 ± 20 150 ± 9 ^b

(0.4mg / kg) (29) (137) (133) (67)

MIC-l + t-PA 4 26 ± 4 ^b 36 ± 3 ^bce 312 ± 38 122 ± 5 ^bcd

(lOrogO + 0.4rag / kg) (11) (189) (151) (55)

0 is X for cost n-A

* p <0.05? s. Kojitsuro-Dunnett) ^d p <0.057 S. MIC-1 (Dunnett) bp 0 / Olvs-co: / Toro-MDunnett) ^β p <0. Olvs. MIC-1 (Dunnett) CP <0.05 vs. t-PA (Dunnett)

A peak time based on the physiological fibrinolysis of the control was observed after about 4 hours.

In the case of animals administered MIC-1 at lOrag / kg / day for 3 days, no significant difference was observed in the control at about 130 minutes. A single administration of t-PA at 0.4 mg / kg after surgery showed a significant difference in control sigma-ru in about 70 minutes. (P <0.01).

 When t-PA was administered to animals to which MIC-1 was administered, a significant difference was observed in the control at about 25 minutes (P <0.01).

 The average control beak height was 19%. The beak height was 22% for animals administered MIC-1 at 10 mg / kg / da for 3 days, and 26% for t-PA, and there was no significant difference in control between them. Was. When t-Δλ was administered to MIC-1 treated animals, a significant difference in control was observed in 36% (P <0.01).

 MIC-1 + t-PA was significantly different from MIC-1 and t-PA in S8 (P <0.01, P <0.05, respectively).

 The average of the total X of dose of the control was 206, 252 when MIC-1 was orally administered at 10 mg / kg / day for 3 days, and 273 when t-PA was administered. Was shown, but no significant difference from the control was observed. When t-PA was administered to animals to which MIC-1 was administered, a significant difference was observed in the control at 312 (p <0.05).

Τ The average value of the control value is 223 minutes for 値₅₀ , and 175 minutes (78% of the control) and t-PA are administered when MIC-1 is administered at 10 rag / kg / day for 3 days. It took 150 minutes (67% of control).

 When t-PA was administered to the animal to which MIC-1 was administered, the time was 122 minutes (55% of the control). In all of these groups, there was a significant difference in the control (MIC 0,05, t-PA, MIC-1 + when MIC-1 was administered at 10 mg / kg / day for 3 days). t-PA is less than P 0.01). MIC-1 + t-PA showed a significant difference between IC-1 and t-PMi: SS (P <0.05).

T _s for MiC-1 alone. Only 值 showed significant drug efficacy against the control. Vehicle and T _{s for} t-PA alone. In planting Control showed significant drug efficacy. MIC-1 and t-PA the combination with If beak time, Bikuhai door, the sum of the X of dose billion, and Τ _5. In the case of Mic-1 alone or t-PA alone, beak height and T _s were significant. In 镲, significant drug efficacy was observed.

 The above results revealed that MIC-l (10fflg / kg) had a thrombolysis promoting effect. When MI C-1 and t-PA were used in combination, the effect was remarkable as compared with the case where each was used alone. It is clear that the law will enhance the therapeutic effect.

 Toxicity

 When MI C-1 was administered to five mice at 1,000 rag / kg 柽 mouth,

No mice died in M.

 When MI C-1 was administered to five rats at a dose of 1,000 mg / kg, no rat died at 72 hours. It is clear that the acute toxicity of the present invention is very low. Formulation Example 1

 — 锭 (150 mg) per 100 mg of the compound of the present invention (MIC-1) lOOrag, lactose 25 tng, corn starch 10 rng, low-substitution hydroxypropyl cell mouth 7.5 mg, hydroxybile mouth cell 2. Tablets were prepared according to a conventional method so as to contain 5 mg of 5 nig of magnesium stearate. Formulation Example 2

According to a conventional method, the compound of the present invention (MlC-l) lOOfflg and sodium chloride 45oig are contained per tube (5 ml) using a distillation column for injection. An injection was prepared.

 Simple sharps of the drawing

 FIG. 1 shows a fibrin dissolution curve obtained by adding 1 ml of UK (30 U / ral) to a fibrin clot obtained from plasma obtained 72 hours after administration of MIC-1 and obtained in Test Example 1.

 The horizontal axis represents the incubation time (hours), and the vertical axis represents the dissolution rate (%). ◊ indicates MIC-1 at 30 mg / kg, MM indicates MIC-1 at lOmg / kg, mouth indicates MIC-1 at 3 mg / kg, and 〇 indicates control. .

 Figure 2 shows the dissolution curve of the fibrin mass obtained by adding 1 ml of UK (30 U / ral) to the fibrin mass produced from plasma obtained 54 hours after MDR-14 administration obtained in Test Example 1. Show.

 The horizontal axis represents the time of incubation (time), and the vertical axis represents the dissolution rate (%). □ indicates the control when 100 tng / kg of MDR-14 was administered, and 〇 indicates the control.

Claims

The scope of the claims (1) A pharmaceutical composition for treating thrombosis, comprising 1-deoxymanno jilimycin or a pharmacologically acceptable acid addition salt thereof as a main component.