RU2290195C1 - Agent possessing anti-coagulating, anti-thrombocytic, anti-thrombosis, fibrin-depolymerizing and fibrinolytic activity - Google Patents

Abstract

FIELD: medicine, chemical-pharmaceutical industry.

SUBSTANCE: invention relates to an anti-coagulating, anti-thrombocytic, anti-thrombosis, fibrin-depolymerizing and fibrinolytic agent containing heptapeptide selank of the formula: Thr-Lys-Pro-Arg-Pro-Gly-Pro. The claimed agent possesses high activity and effectiveness as compared with the known agents.

EFFECT: enhanced medicinal properties of agent.

2 tbl, 3 ex

Description

The invention relates to medicine, to agents having anticoagulant, fibrindepolymerization, antiplatelet, antithrombotic and fibrinolytic activity.

Among anticoagulant pharmaceuticals, heparin is known to have anticoagulant activity (Zaman SM, Neiring P., Gandhi MR, Gaines PA The pharmacokinetics and US usage of heparin in vascular intervention. // Clin. Radiol. 1996. V 51. No. 2. P. 113-116.) [1].

However, it has the following disadvantages:

1) does not have antithrombotic properties,

2) does not have fibrinolytic effect,

3) in case of an overdose, high molecular fractions can cause undesirable side effects of bleeding.

A known preparation of acetylsalicylic acid, which has an antiplatelet effect (McKee SA, Sane DC, Deliargyris EN Aspirin resistance in cardiovascular disease: a review of prevalence mechanisms, and clinical significance. // Thromb. Haemost. (Schattauer). 2002. V.88. No. 5. P.711-715) [2].

The drug has the following disadvantages:

1) does not detect anticoagulant activity,

2) has a weak fibrindepolymerization effect,

3) has side effects - bleeding from the gastrointestinal tract.

A known plasmin preparation having a fibrinolytic effect (Gitabert J., Estelles A., Grancella S., Espana F., Aznar J. Fibrinolytic system and reproductive process with special reference to fibrinolytic failure in preeclampsia. // Hum. Reprod. 1995. V .10. Suppl. 2. P.121-131.) [3].

It has the following disadvantages:

1) does not have anticoagulant effects,

2) does not have antiplatelet action,

3) does not have fibrinopolymerization activity,

4) exhibits a proteolytic effect.

Peptides, fibrinogen cleavage products with antithrombotic, antiplatelet activity and causing local fibrinolysis (Standker L., Sillard R., Bensch KW, Ruf A., Raida M., Schulz-Knappe P., Shenky AG, Patscheke H., Forssmann W.-G, In vivo degradation of human fibrinogen Aα: detection of cleavage sites and release of antithrombotic peptides. // Biochem. Biophys. Research communications. 1995. V.215. No. 3. P. 826-832.) [4 ].

However, these peptides did not show anticoagulant, fibrin-depolymerization and general fibrinolytic activity.

Known heptapetide of the formula Thr-Lys-Pro-Arg-Pro-Gly-Pro, called selank, with anti-anxiety effect (Seredenin SB, Kozlovskaya MM, Blednev Yu.A. et al. Study of the anti-anxiety action of an endogenous analogue tafcin peptide in inbred mice with different phenotypes of emotional stress reactions. // GNI Journal. 1998. V. 48. Issue 1.) [5].

However, the anticoagulant, fibrindepolymerization and fibrinolytic properties of this known heptapeptide are not described.

The technical result achieved by the present invention is the identification and recommendation of an agent combining anticoagulant, antiplatelet, antithrombotic, fibrindepolymerization and fibrinolytic activity.

The specified technical result is achieved by the fact that the selank heptapeptide of the formula: Thr-Lys-Pro-Arg-Pro-Gly-Pro is used as an agent with anticoagulant, antiplatelet and antithrombotic, fibrindepolymerization, fibrinolytic activity.

The following are examples illustrating the invention.

Example 1

Take 1 mg of the Selank heptapeptide preparation - Thr-Lys-Pro-Arg-Pro-Gly-Pro, dissolved in 2.5 ml of physiological NaCl solution and injected intravenously into 5 laboratory rats of 0.5 ml (200 mcg) in v.jugularis ) per 200 g of body weight. A control animal was injected with a 0.85% NaCl solution in a dose of 0.5 ml per 200 g of body weight in the same way. After 10 minutes, blood is drawn and centrifuged at 1000 rpm for 5 minutes to obtain platelet-rich plasma (for platelet aggregation analysis), then blood is further centrifuged for 10 minutes at 2000 rpm to obtain platelet-poor plasma. In platelet-rich plasma, platelet aggregation in experimental samples was 68.8%, while in control it was 100%, i.e. Selank heptapeptide has high antiplatelet activity. In platelet-poor plasma, an increase in enzymatic fibrinolytic activity was detected due to an increase in plasminogen activator activity by 2.3 times, total fibrinolytic activity - by 2.1 times, anticoagulant activity - by 1.2 times, fibrin-depolymerization activity - by 8 times compared with the control (saline NaCl).

Example 2

Take 1 mg of the drug Selank, dissolved in 5 ml of saline. Next, proceed as described in example No. 1.

The analysis of the parameters of hemostasis reveals the following.

In platelet-rich plasma, platelet aggregation in experimental samples was 83%, while in control it was 100%, i.e. heptapeptide has antiplatelet activity. In platelet-poor plasma, an increase in enzymatic fibrinolytic activity was detected due to increased plasminogen activator activity by 1.7 times, total fibrinolytic activity - 1.6 times, anticoagulant activity - 1.4 times, fibrin-desolimerization activity - 1.8 times compared with control (saline).

Example 3

Take 1 mg of the preparation Thr-Lys-Pro-Arg-Pro-Gly-Pro selank peptide, dissolved in 2.5 ml of physiological NaCl and administered intranasally for 5 days every 24 hours to 5 laboratory 0.5 ml rats per 200 g of body weight. Control animals in the same way injected 0.85% NaCl solution. 5 days after the last injection, blood was taken from v.jugularis and centrifuged at 2000 rpm for 10 min to obtain platelet-poor plasma. Then, in the v.jugularis region (stasis) isolated by clamps, thrombin is caused by thrombin formation and the antithrombotic effect of the drug is determined after 1 hour.

In a platelet-poor plasma, an increase in total fibricolytic activity is detected - 2 times due to an increase in the activity of the plasminogen activator, anticoagulant activity - 1.5 times, fibrin-depolymerization activity - 2 times compared with the control (saline). The drug Selank exhibited a high antithrombotic effect that exceeded the antithrombotic properties of the blood plasma of healthy rats by 5.6 times: while in the control the blood clot weight was 1.4 mg, in the experiment it was only 0.27 mg.

Table 1 shows comparative data of the proposed drug Selank and known [4].

Table 1. Properties of target products Famous [4] The proposed peptide is Selank Activities 1) anticoagulant Are absent Are available 2) enzymatic fibrinolysis in the bloodstream Local General 3) fibrindepolymerization, fibrinolytic Absent Is available 4) antiplatelet Is available Is available 5) antithrombotic Is available Is available

As can be seen from table 1, the proposed drug Selank, in contrast to the known [4], in vitro has additional combined positive properties - anticoagulant, fibrindepolymerization, enhancing the total fibrinolytic activity in blood plasma. In contrast to the known peptides [4], the proposed preparation Selank, when it appears in animals, has both anticoagulant, fibrinolytic, antiplatelet, antithrombotic and fibrindepolymerization effects.

Thus, the well-known heptapeptide Selank combines anticoagulant, fibrindepolymerization, antiplatelet, antithrombotic and fibrinolytic properties.

Table 2 presents data demonstrating the advantages of the claimed Selank peptide Thr-Lys-Pro-Arg-Pro-Gly-Pro in comparison with the reference preparations of the heparin-fibrinogen complex and tri- and tetrapeptides (Pro-Gly-Pro and Gly-Pro- Gly-Gly).

table 2 Activities Selank Thr-Lys-Pro-Arg-Pro-Gly-Pro Fibrinogengeparin complex (prototype) Pro-Gly-Pro (prototype) Gly-Pro-Gly-Gly (prototype) In vitro Anticoagulant 120-143% 140% 122-140% 118% Fibrinolytic (total) 36-42 mm ² 30-36 mm ² 36-42 mm ² 24 mm ² Antiplatelet 61-68% - 83% 75% In vivo Intravenous administration Anticoagulant activity 121% 135% 173% one hundred% Fibrinolytic non-enzymatic activity 150% 140% 173% one hundred% Tissue plasminogen activator activity 298% 103% 220% one hundred% Platelet Aggregation 68.8% - 83% one hundred% Intranasal administration Antithrombotic activity (by weight of blood clots) 19% 48% 19% one hundred%

Thus, from table 2 it is seen that the claimed Selank peptide (Thr-Lys-Pro-Arg-Pro-Gly-Pro) has advantages over the Pro-Gly-Pro and Gly-Pro-Gly-Gly peptides in terms of the increase in anticoagulant, fibrinolytic and antiplatelet activity in vitro: when administered intravenously, the anticoagulant-fibrinolytic effects of the claimed peptide exceed the same activities of the reference Gly-Pro-Gly-Gly tetrapeptide; upon intranasal administration, the antithrombotic effects of Selank (Thr-Lys-Pro-Arg-Pro-Gly-Pro) exceed the similar effect of the fibrinogen-heparin complex and the Gly-Pro-Gly-Gly tetrapeptide and correspond to the Pro-Gly-Pro effect.

Claims (1)

The use of heptapeptide selank of the formula Thr-Lys-Pro-Arg-Pro-Gly-Pro as an agent with anticoagulant, antiplatelet, antithrombotic, fibrindepolymerization and fibrinolytic activities.