Use of fibrinogen in preparing medicine for preventing and treating aortic dissection

Abstract

The present application belongs to the technical field of pharmaceutical preparations, more specifically to the technical field of cardiovascular disease therapeutic drugs, and particularly relates to a new use of fibrinogen for preparing drugs for preventing and treating aortic dissection. The present invention, through the study of the pharmacological effects of fibrinogen, discovered and verified the use of fibrinogen in the treatment of acute aortic dissection. Fibrinogen can reduce the breakage of elastic fibers in aortic tissue and the degradation of elastin, and maintain the contractile phenotype of VSMC, thereby effectively reducing the dissection rupture rate and the mortality rate of patients with aortic dissection. It plays a significant role in improving acute active dissection, filling the gap in drug treatment measures for acute aortic dissection.

Description

Technical Field

[0001] The present application belongs to the technical field of pharmaceutical preparations, more specifically to the technical field of cardiovascular disease therapeutic drugs, and particularly relates to a new use of fibrinogen for preparing drugs for preventing and treating aortic dissection. Background Art

[0002] Aortic dissection (AD) refers to the separation of the intima and media caused by damage to the aortic wall, with blood flowing in the true and false cavities. According to the location of accumulation, it can be divided into Stanford type A aortic dissection (STAAD) and type B aortic dissection. In my country, 3 out of every 100,000 people suffer from aortic dissection, especially acute STAAD, which is extremely dangerous. It is an extremely dangerous cardiovascular emergency that seriously threatens human life and health.

[0003] At present, medical guidelines recommend surgical treatment as the first choice for the treatment of STAAD. However, surgical treatment is not only risky, difficult, and expensive, but also some patients are unable to receive surgical treatment for various reasons and can only choose conservative drug treatment. However, existing drug treatments are only symptomatic treatments for lowering blood pressure, controlling heart rate, and relieving pain, and cannot repair the aorta and reduce rupture rates and mortality. Therefore, this field urgently needs to develop targeted intervention drugs that participate in STAAD protection to stabilize patients' conditions, reduce rupture rates, and reduce complications and mortality.

[0004] Current studies believe that the pathogenesis of STAAD includes phenotypic transformation of vascular smooth muscle cells (VSMCs), degradation of extracellular matrix, inflammation, oxidative stress, mitochondrial dysfunction or epigenetics. The aorta is composed of the intima, media and adventitia layers. VSMCs are the main cellular component of the aortic media and play a core role in maintaining aortic function and homeostasis. According to the functional characteristics of VSMCs, they can be divided into contractile (differentiated) and proliferative (undifferentiated) types. The main function of contractile VSMCs is to contract blood vessels and regulate vascular tension, blood pressure and blood flow distribution, while their proliferation, migration and matrix synthesis abilities are poorer than those of proliferative VSMCs, and they are mature types of VSMCs. In contrast to contractile VSMCs, proliferative VSMCs have no contractile function and mainly synthesize and secrete extracellular matrix, such as collagen and matrix metalloproteinases. Their differentiation is low or even undifferentiated.

[0005] Existing studies have shown that under certain stimuli, VSMCs can transform from a differentiated contractile state to a proliferative state, which is characterized by a decrease in the expression of contractile markers and an increase in proliferation and secretion markers. The contractile activity of VSMCs is the basis of aortic structure and function. Under normal physiological conditions, most VSMCs in the ascending aorta wall have a contractile phenotype. The phenotype that transforms to a proliferative state is considered to be the main driving factor of aortic dissection. Reversing the phenotypic transformation of VSMCs can participate in the repair of STAAD and play a therapeutic role.

[0006] Fibrinogen (FIB) is a protein with coagulation function that is mainly synthesized by hepatocytes. It is the coagulation factor with the highest content in plasma. Its most basic physiological function is to participate in the final stage of the coagulation process and convert into fibrin to form a blood clot. It is often used in clinical practice to stop bleeding. In addition, FIB can also participate in matrix physiology and is closely related to the regulation of inflammatory processes, infection, wound healing, cell-cell interactions, cell migration, tumor growth and angiogenesis. At present, there are no reports on the protective effect of FIB on STAAD. Developing it into a therapeutic drug for acute aortic dissection has extremely high potential value and social significance. Summary of the invention

[0007] Therefore, the technical problem to be solved by the present invention is to provide a new use of fibrinogen for preparing a drug for preventing and treating aortic dissection;

[0008] The second technical problem to be solved by the present invention is to provide a pharmaceutical preparation based on fibrinogen as an active ingredient for preventing and treating aortic dissection.

[0009] In order to solve the above technical problems, the present invention provides a use of fibrinogen for preparing a drug for preventing and treating aortic dissection.

[0010] Specifically, the fibrinogen is used as the only active ingredient.

[0011] Specifically, the aortic dissection includes Stanford type A aortic dissection.

[0012] Specifically, the aortic dissection includes acute aortic dissection.

[0013] Specifically, the aortic dissection includes acute Stanford type A aortic dissection.

[0014] Specifically, the effective dosage of fibrinogen is 350-400 mg / kg body weight, administered once every three days, preferably injected into the tail vein once every three days.

[0015] The invention also discloses a pharmaceutical preparation for preventing and treating aortic dissection, wherein the active ingredient of the pharmaceutical preparation includes fibrinogen.

[0016] Specifically, the pharmaceutical preparation for preventing and treating aortic dissection includes a clinically acceptable oral preparation or injection preparation.

[0017] Preferably, the pharmaceutical preparation is a clinically acceptable injection preparation.

[0018] Specifically, the pharmaceutical preparation for preventing and treating aortic dissection, the injection preparation includes an injection solution, a sterile powder for injection or a concentrated solution for injection, preferably a sterile powder for injection.

[0019] The so-called injection solution, sterile powder for injection and concentrated solution for injection are the classification methods of injection dosage forms specified in the pharmacopoeia. The commonly used dosage form definitions such as water injection and powder injection in pharmacy are also included.

[0020] Preferably, the fibrinogen is used for patients with acute aortic dissection to protect them from rupture and death, and is administered by intravenous injection, under the condition that it is given immediately after the dissection patient is admitted to the hospital.

[0021] Specifically, the pharmaceutical preparation for preventing and treating aortic dissection comprises pharmaceutically acceptable excipients.

[0022] Specifically, in the pharmaceutical preparation for preventing and treating aortic dissection, the auxiliary materials include one or more of excipients, solubilizers, stabilizers, antioxidants, pH regulators or osmotic pressure regulators.

[0023] The present invention also discloses a method for preparing the pharmaceutical preparation for preventing and treating aortic dissection, comprising the steps of taking the fibrinogen as an active ingredient and adding selected auxiliary materials to prepare a clinically acceptable pharmaceutical preparation.

[0024] The present invention also discloses a use of fibrinogen for preparing a drug having at least one of the following effects (a)-(f):

[0025] (a) Reduce the rate of aortic clamp rupture;

[0026] (b) reduce the mortality rate of patients with aortic dissection;

[0027] (c) reduce the breakage of elastic fibers and the degradation of elastin in aortic tissue;

[0028] (d) maintain the contractile phenotype of VSMCs;

[0029] (e) Repairing damaged aortic wall;

[0030] (f) Protect vascular smooth muscle cells.

[0031] Specifically, the fibrinogen is used as the only active ingredient.

[0032] The present invention has discovered and verified the use of fibrinogen in treating acute aortic dissection through the study of the pharmacological effects of fibrinogen. Fibrinogen can reduce the breakage of elastic fibers in aortic tissue and the degradation of elastin, and maintain the contractile phenotype of VSMC, thereby effectively reducing the rupture rate of dissection and the mortality rate of patients with aortic dissection. It plays a significant role in improving acute active dissection and fills the gap in drug treatment measures for acute aortic dissection.

[0033] The present invention verifies for the first time the mechanism of fibrinogen in treating aortic dissection through the study of the pharmacological effects of fibrinogen, and further provides the use of fibrinogen in the preparation of drugs for treating aortic dissection, providing a solid pharmacodynamic basis for new clinical research and application of the drug, and has broad market prospects after system development. It also provides an example for the medicinal use of classic hemostatic drugs and the "new use of old drugs", which can be applied more quickly in clinical practice. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to make the content of the present invention more clearly understood, the present invention is further described in detail below according to specific embodiments of the present invention in conjunction with the accompanying drawings, wherein:

[0035] Figure 1 The results of constructing the mouse STAAD model in Example 1 and the protective effect of FIB in the model; A: The ultrasound results of small animals after BAPN feeding for 4 weeks showed that the aortic arch of mice was significantly dilated; B: Ultrasound statistical graph; C: Kaplan-Meier survival curves of the STAAD model of mice in different treatment groups; D: Gross images of mice in different treatment groups after sampling;

[0036] Figure 2 Staining of mouse aortic wall tissue sections; A: H&E staining; B: EVG staining;

[0037] Figure 3 The immunofluorescence staining results of the mouse aorta wall in Example 2; wherein, A: immunofluorescence staining of type III collagen and elastic fibers in the mouse aorta wall; B: statistical graph of type III collagen;

[0038] Figure 4 The results of immunofluorescence staining of mouse aorta in Example 3; wherein, A: immunofluorescence staining of proliferation marker OPN and contraction marker α-SMA in aorta of mice in different treatment groups; B: OPN statistical graph; C: α-SMA statistical graph;

[0039] Figure 5The expression of marker proteins of phenotypic transformation of mouse aorta tissue after FIB treatment detected by Western blot in Example 4 and the statistical graph;

[0040] Figure 6 These are the staining results of F-actin and G-actin after human primary VSMCs were treated with FIB in Example 5. DETAILED DESCRIPTION

[0041] The present invention is further described in detail below in conjunction with the embodiments. The technical solutions in the embodiments are only for exemplary display and do not constitute any limitation on the protection scope of the present application, which is defined by the claims.

[0042] Fibrinogen is a protein with coagulation function mainly synthesized by liver cells. It is the coagulation factor with the highest content in plasma. The molecular weight of fibrinogen is about 340kDa. It is in the shape of a tripartite sphere and consists of three pairs of different polypeptide chains: α, β, and γ. The two ends are D regions and the middle is E region. The D region and the E region are connected by three α-helical peptide chains.

[0043] Fibrinogen is an important reaction substrate for thrombosis and participates in the key steps of thrombosis. 2+ Under the action of coagulation factors such as TNF-α and TNF-α, fibrin monomers are formed and covalently bound to each other to form fibrin polymers. The α chains are staggered and overlapped covalently cross-linked to form a stable fibrin network, which eventually entrains red blood cells (RBC), platelets (PLT), α2-antiplasmin (α2-AP) and other components to form a stable thrombus structure.

[0044] Currently, the clinical uses of fibrinogen include: congenital fibrinogen deficiency or hypofibrinogenemia; or acquired fibrinogenemia, such as severe liver damage, cirrhosis, disseminated intravascular coagulation, postpartum hemorrhage, and coagulation disorders caused by fibrinogen deficiency due to major surgery, trauma or internal bleeding.

[0045] Clinically, the clinical dosage form of the fibrinogen drug is an injection dosage form, which is usually a grayish white or light yellow loose body. After reconstitution, the solution is clear or slightly opalescent, and a small amount of fine flocs is allowed.

[0046] Clinically, the fibrinogen drug is used as follows: preheat the product and sterile water for injection to 30-37°C before use, then inject the preheated sterile water for injection according to the amount indicated on the bottle label, place in a 30-37°C water bath, and gently shake to dissolve the product completely (avoid violent shaking to avoid protein denaturation). Use an infusion set with a filter device for intravenous drip. The drip rate is generally about 60 drops per minute. The dosage of the fibrinogen drug should be determined according to the condition and clinical test results. Generally, 1-2g is given for the first time. If necessary, continue to administer the drug according to the doctor's advice.

[0047] It should be noted that in the following embodiments of the present invention, fibrinogen can be used interchangeably with Fibrinogen and FIB to represent the same meaning.

[0048] Example 1 Construction of STAAD model and protective effect of FIB in the model

[0049] In this example, a mouse STAAD model was constructed. The BAPN modeling group (n=20) used 3-week-old C57BL / 6 background wild-type male mice and fed with a feed containing 0.4% β-aminopropionitrile (BAPN) for 4 weeks, and the control group (n=20) was fed with a normal diet; the FIB treatment group (n=20) was injected with FIB through the tail vein of mice after 2 weeks of BAPN feeding (injected once every 3 days, each dose of 375 mg / kg, weighed before each injection of the drug, and the drug dosage was adjusted) for 2 weeks. The dead mice were immediately dissected to determine whether they died due to TAAD rupture, and serum and aortic wall tissue were collected and the survival curve was recorded.

[0050] In this embodiment, according to the above operation, an aortic ultrasound examination was performed after 4 weeks of feeding. Figure 1 As shown in Figure AB, the aorta of BAPN-fed mice showed obvious dilation and dissection. Figure 1 As shown in Figure C, at 2 weeks, some mice in the BAPN group began to die from dissection, but after FIB treatment, the dissection rupture and mortality of mice were significantly reduced. Figure 1 As shown in D, the gross anatomical diagram further demonstrates the modeling results and the protective effect after FIB treatment. The results of this example show that FIB can play a protective role in mouse STAAD.

[0051] Example 2 FIB reduces the effect of elastic fiber rupture in aortic tissue

[0052] The frozen sections of mouse aorta tissue were obtained by the same method as in Example 1 and subjected to H&E staining and EVG staining. Figure 2 As shown in Figure 2, FIB treatment can significantly improve aortic lesions caused by BAPN (such as Figure 2Middle A), relieve the rupture of elastic fibers caused by BAPN (such as Figure 2 (middle B).

[0053] This example further performed immunofluorescence staining of type III collagen (elastin) on mouse aorta tissue. The results are shown in the attached Figure 3 As shown in the figure, after 4 weeks of BAPN feeding, the expression of type III collagen was significantly reduced, accompanied by obvious rupture of elastic fibers. After 2 weeks of FIB treatment, the expression of type III collagen returned to normal levels, and the rupture of elastic fibers was significantly alleviated (as shown in the figure). Figure 3 (middle AB).

[0054] Example 3 FIB can maintain the contractile phenotype of VSMC

[0055] The mice were treated in the same manner as in Example 1. The STAAD model of mice was established by feeding with BAPN. After 2 weeks of BAPN feeding, the mice were treated with FIB. After 2 weeks of FIB treatment, the mice were killed and the aortic tissues of the mice were obtained and immunofluorescence staining was performed. The results are shown in the attached figure. Figure 4 shown.

[0056] Figure 4 The results shown in AC indicate that after 4 weeks of BAPN feeding, there was no significant change in α-SMA, a contraction phenotype marker of the mouse aortic wall; however, after 2 weeks of BAPN feeding and concurrent FIB treatment, OPN expression was significantly decreased, while α-SMA expression was significantly increased, indicating that FIB treatment can transform mouse aortic VSMCs from a proliferative type to a contractile type.

[0057] Example 4

[0058] This example further used the above mouse aorta tissue protein and used Western blot to detect the changes in phenotypic transformation markers after FIB treatment. The results are shown in the attached Figure 5 shown.

[0059] The above results indicate that FIBs can maintain the contractile phenotype of VSMCs.

[0060] Example 5

[0061] This example further conducted an in vitro experiment using human primary VSMC cells, treated with FIB (3 mg / ml), and then stained for F-actin and G-actin. The results are shown in the attached Figure 6 shown.

[0062] The above results indicate that FIB treatment can promote the assembly of F-actin and reduce its dissociation into G-actin, further indicating that FIB treatment can maintain the contractile phenotype of VSMCs.

[0063] Example 6

[0064] The drug for preventing and treating aortic dissection described in this embodiment contains fibrinogen as an active ingredient, or fibrinogen can be selected as the only active ingredient.

[0065] The drug of the present invention may also contain other Chinese and Western medicines or health products that have therapeutic or auxiliary therapeutic effects on aortic dissection, or the drug of the present invention may be used in combination with these Chinese and Western medicines or health products or surgical treatments. The Chinese and Western medicines or health products include but are not limited to antihypertensive drugs, analgesics, heart rate control drugs, etc.

[0066] The pharmaceutical preparations used in the present invention can be selected from the types of preparations suitable for oral administration or injection administration. For those skilled in the art, the following dosage forms can contain fibrinogen as an active ingredient to prepare the desired pharmaceutical preparations.

[0067] As a clinically applicable product, the drug of the present invention is preferably formulated into a preparation for parenteral administration (e.g., injection, such as bolus injection or continuous infusion), and can be present in the form of a unit dose in an ampoule, a prefilled syringe, a small volume infusion bag or a multi-dose container together with an added preservative. The drug can be in the form of a suspension, solution or emulsion of an oily or aqueous carrier, and can contain formulation ingredients such as suspending agents, stabilizers and / or dispersants. In addition, the active ingredient can be in the form of a powder, which can be obtained by aseptic separation of a sterilized solid or by freeze-drying of a solution, for reconstitution with a suitable carrier such as sterile, pyrogen-free water before use.

[0068] In the therapeutic use of the pharmaceutical preparation, the daily dosage of the fibrinogen can be in accordance with conventional dosages. These dosages can vary according to the needs of the patient, the severity of the disease to be treated, and the drug used. Generally speaking, treatment is started with a smaller dosage than the optimal dosage of the drug, and thereafter, the dosage is increased in small amounts to achieve the best effect. For convenience, the total daily dosage can be further divided into divided doses within a day if necessary.

[0069] In summary, the fibrinogen described in the present invention is used for patients with acute aortic dissection to protect them from rupture and death. The preferred method of use is intravenous injection, and the condition of use is to give it to patients with dissection immediately after admission to the hospital.

[0070] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A use of fibrinogen for preparing a drug for treating Stanford type A aortic dissection induced by BAPN; The therapeutic uses of the fibrinogen include: (a) Reduce the rate of aortic clamp rupture; (b) reduce the mortality rate of patients with aortic dissection; (c) reduce the breakage of elastic fibers and the degradation of elastin in aortic tissue; (d) maintain the contractile phenotype of VSMCs; (e) Repair of damaged aortic wall; (f) Protect vascular smooth muscle cells.

2. The use according to claim 1, characterized in that The fibrinogen is used as the only active ingredient.

3. The use according to claim 2, characterized in that: The effective amount of fibrinogen is 350-400 mg / kg body weight.

4. The use according to any one of claims 1 to 3, characterized in that: The medicine includes a clinically acceptable oral preparation or injection preparation.

5. The use according to claim 4, characterized in that The injection preparation includes injection solution, sterile powder for injection or concentrated solution for injection.

6. The use according to any one of claims 1 to 3, characterized in that: The drug includes pharmaceutically acceptable excipients.

7. The use according to claim 6, characterized in that The auxiliary materials include one or more of excipients, solubilizers, stabilizers, antioxidants, pH regulators or osmotic pressure regulators.

8. The use according to any one of claims 1 to 3, characterized in that: The preparation method of the drug comprises the steps of taking the fibrinogen as an active ingredient and adding selected auxiliary materials to prepare a clinically acceptable drug preparation.

Images