Use of pirfenidone in the manufacture of a medicament for the treatment of deep vein thrombosis

Abstract

The application provides application of pirfenidone in preparation of a medicine for treating deep vein thrombosis, wherein the structure of the pirfenidone is shown in the following formula (I): It is proved by experiments that the pirfenidone has good effects on deep vein thrombosis, has no adverse reactions, can slow down deep vein thrombosis of a rat induced by subtotal ligation of an inferior vena cava, and has a good application prospect in treatment, alleviation or improvement of deep vein thrombosis.

Description

Technical Field

[0001] This invention relates to a novel use of pirfenidone, specifically, to the use of pirfenidone in the treatment of deep vein thrombosis. Background Technology

[0002] Deep venous thrombosis (DVT) is a disease caused by abnormal blood clotting in deep veins, leading to impaired venous return. It commonly occurs in the lower extremities, is clinically prevalent, and can be fatal. Globally, DVT has a high incidence rate, which is increasing annually. Clinical studies report a mortality rate of approximately 15% within the first three months after diagnosis. DVT is divided into acute, subacute, and chronic phases, with early DVT encompassing both acute and subacute phases. Early lower extremity DVT primarily manifests as sudden swelling and pain in the affected limb. Severe cases may present with phlegm palpebrarum, the most serious form of lower extremity DVT. This is due to thrombosis of the iliofemoral vein and its tributaries, severely obstructing venous return, causing extremely high tissue tension, leading to compression and spasm of the lower extremity arteries, and limb ischemia. Clinically, this presents as extreme swelling and severe pain in the lower extremities, a shiny, bluish-purple skin, and the disappearance of the dorsalis pedis artery pulsation. If not treated promptly, shock and venous gangrene can occur. In the chronic phase, post-thrombotic syndrome (PTS) can develop due to chronic fibrosis caused by thrombosis. This syndrome manifests as clinical signs of chronic lower extremity venous insufficiency, including heaviness, swelling, pain, varicose veins, pruritus, pigmentation, and eczema in the affected limb. In severe cases, it can lead to extensive lower extremity swelling, lipotic scleroderma, and persistent ulcers, significantly impacting the patient's quality of life. However, its pathogenesis remains unclear, but it is generally believed to be caused by the interaction of many factors, including genetic, infectious, and immune factors.

[0003] Traditional first-line treatments for deep vein thrombosis (DVT) are limited and singular, primarily consisting of anticoagulants such as low molecular weight heparin, warfarin, and rivaroxaban. While these drugs effectively prevent the formation of fresh thrombi, they cannot dissolve existing DVT thrombi. Undissolved thrombi can further develop into chronic fibrosis, eventually forming the chronic phase of DVT. The insolubility of thrombi in chronic fibrosis promotes the formation of new thrombi, increasing the risk of long-term thrombus detachment and pulmonary embolism (PTS), which often leads to sudden death. Even with adequate anticoagulation, approximately 25% to 50% of DVT patients develop PTS due to persistent thrombi and chronic fibrosis. Therefore, current anticoagulant therapy remains ineffective and carries serious side effects such as bleeding risks. Other treatment options remain controversial, have numerous clinical limitations, and pose significant risks of side effects, potentially severely impacting patients' quality of life long-term, making them unacceptable to many. Therefore, there is an urgent need for candidate drugs extracted from natural foods and plants for DVT treatment, such as flavonoids, polyphenols, terpenes, and alkaloids.

[0004] Pirfenidone (PFD) is a drug approved in China for the treatment of human idiopathic pulmonary interstitial fibrosis. PFD has been shown to primarily act on potentially influential growth factor receptors in the pathophysiological mechanisms of pulmonary fibrosis, most importantly transforming growth factor receptor (TGF-β1R) and the major pro-fibrotic signaling pathway Wnt / β-catenin. Patients with deep vein thrombosis (DVT) often present with undissolved thrombi and chronic fibrosis. However, to date, there are no reports of PFD being effective in treating DVT. Summary of the Invention

[0005] In view of this, the present invention aims to propose a new use of pirfenidone, namely, the use of pirfenidone in the preparation of a medicament for treating DVT, wherein the structure of said pirfenidone is shown in the following formula (I):

[0006] In the above applications, the dosage of pirfenidone is 30 mg / kg to 300 mg / kg.

[0007] The present invention also provides a medicament for treating DVT, comprising pirfenidone, and pharmaceutically acceptable salts, esters, hydrates, and excipients of pirfenidone.

[0008] Among the above-mentioned drugs, the pirfenidone salts that are pharmaceutically acceptable include both organic and inorganic salts.

[0009] In the above-mentioned drugs, the organic salts include one or more of the following: methanesulfonate, formate, acetate, trifluoroacetate, maleate, tartrate, succinate, fumarate, citrate, benzenesulfonate, p-toluenesulfonate, naphthalenesulfonate, lactate, and benzoate; and the inorganic salts include one or more of the following: hydrochloride, hydrobromide, sulfate, and phosphate.

[0010] Among the above-mentioned drugs, the dosage form is selected from tablets, capsules, pills, suppositories, aerosols, oral liquid preparations, granules, powders, injections, syrups, tinctures, lotions, and films.

[0011] Among the above-mentioned drugs, the routes of administration include oral, injection, implantation, external application, spray, inhalation, or combinations thereof.

[0012] Compared with the prior art, the present invention has the following advantages: The study of this invention shows that pirfenidone has good efficacy against DVT with no adverse reactions, and can alleviate DVT in rats with subtotal ligation of the inferior vena cava. It has good application prospects in the treatment, relief or improvement of DVT disease. Attached Figure Description

[0013] Figure 1 The study observed the incidence of thrombosis in rats of different groups over 7 days. IVC represents a rat DVT model with subtotal ligation of the inferior vena cava.

[0014] Figure 2 The study observed the incidence of thrombosis in rats of different groups over 14 days. IVC represents a rat DVT model with subtotal ligation of the inferior vena cava.

[0015] Figure 3 The length of thrombi in rats in different observation periods is shown.

[0016] Figure 4 The values ​​represent the weight of thrombi in rats at different observation times.

[0017] Figure 5 HE staining of thrombus tissue from rats in different observation periods. Figure A: 7-day IVC rats; Figure B: 7-day IVC+PFD rats; Figure C: 14-day IVC rats; Figure D: 14-day IVC+PFD rats. The inferior vena cava contained thrombus tissue (Figures AD, indicated by red arrows), with minimal cellular atypia, no obvious nucleoli, and a homogeneous, wavy, red-stained interstitium consistent with collagen fiber characteristics (Figures A and C, indicated by black arrows). Compared to the IVC group, the IVC+PFD group showed more readily dissolved and shrunk thrombi at the same observation period, with a lighter homogeneous red-stained tissue structure and fewer irregularly shaped cells and oval-shaped nuclei.

[0018] Figure 6 Masson staining of thrombus tissue from rats in different observation periods. Figure A: 7-day IVC rats; Figure B: 7-day IVC+PFD rats; Figure C: 14-day IVC rats; Figure D: 14-day IVC+PFD rats. The thrombus tissue in the inferior vena cava (shown by black arrows in Figures AD) shows blue, wavy tissue in the interstitial space, consistent with the characteristics of collagen fibers (Figures A and C, shown by green arrows). This change is more pronounced in the IVC group with prolonged observation time. Compared to the IVC group, the IVC+PFD group showed more readily dissolved and shrunken thrombi at the same observation time, with a greater reduction in the area of ​​blue positive collagen fibers. Detailed Implementation

[0019] Unless otherwise defined, the technical terms used in the following embodiments have the same meanings as commonly understood by those skilled in the art. Unless otherwise specified, the experimental reagents used in the following embodiments are conventional biochemical reagents; and the experimental methods, unless otherwise specified, are conventional methods. The present invention will be described in detail below with reference to the embodiments.

[0020] This invention provides the use of pirfenidone in the preparation of a medicament for treating deep vein thrombosis, wherein the structure of pirfenidone is shown in formula (I):

[0021] (I) Example

[0022] Effects of pirfenidone on deep vein thrombosis induced by subtotal ligation of the inferior vena cava in rats.

[0023] Experimental Groups: 120 healthy, clean-grade SD rats were randomly divided into three groups: ① sham-operated group (n=40); ② IVC group (n=40); and ③ IVC+PFD group (n=40). In the experimental groups, the inferior vena cava was subtotal ligated to successfully establish the IVC rat model, which is the animal model of deep vein thrombosis (DVT). The sham-operated group underwent only routine laparotomy and suturing. Rats in the IVC+PFD group received daily PFD gavage intervention after model establishment. The three groups were further divided into two subgroups based on observation time: a 7-day group (n=20) and a 14-day group (n=20). All rats were kept in an animal room at a temperature of 20-24℃ and humidity of 65-70%, with free access to food and water.

[0024] Preparation of an animal model of deep vein thrombosis: After fasting for 12 hours before surgery, rats were anesthetized by intraperitoneal injection of ready-to-use tribromoethanol, fixed on a rat board, and the hair on the midline of the rat's abdomen was trimmed. After disinfection with povidone-iodine, a 2cm incision was made in the midline of the abdomen under aseptic conditions to expose and separate the inferior vena cava branches below the left renal vein to the level of the iliac vein. The inferior vena cava branches on both sides were ligated with 5-0 sutures. The abdominal aorta and inferior vena cava were separated about 0.5cm below the junction of the left renal vein and the inferior vena cava. A 4-0 suture was placed parallel to the main trunk of the inferior vena cava, and the 5-0 suture was passed through the inferior vena cava and ligated. The parallel 4-0 sutures were removed. The peritoneum and abdominal muscles were sutured using a continuous suture method, and the outer skin was sutured using a breakpoint suture method. The incision site was wiped with povidone-iodine. After recovery, the rats were able to drink and drink freely.

[0025] Intervention and Tissue Specimen Collection and Preservation: After the establishment of the IVC rat model, the observation period began on the first day after model establishment. Rats in the IVC+PFD group were administered PFD 300 mg / kg by gavage once daily. The IVC group and sham-operated group were administered an equal volume of sterile saline by gavage daily. Subsequent experiments were conducted after 7 and 14 days of continuous gavage. On the 7th and 14th days after model establishment, inferior vena cava thrombus tissue was collected from each group of rats to observe the thrombus incidence, measure thrombus length and weight. The inferior vena cava thrombus tissue was carefully separated using sterile forceps. The thrombus was placed on ice and rinsed thoroughly with sterile saline. The thrombus fluid was absorbed with sterile filter paper, and the thrombus was weighed and its length measured. The proximal portion of the thrombus tissue was cut and placed in a centrifuge tube pre-filled with 4% paraformaldehyde to fix the tissue block. The tissue was then routinely dehydrated, embedded in paraffin, and sectioned for subsequent HE staining, Masson staining, etc. The remaining thrombus tissue was placed in labeled cryovials and stored at -80°C for later use.

[0026] (1) Thrombosis incidence: Observe the thrombosis incidence. After the model is established, use ultrasound to confirm thrombosis and then use pirfenidone for intervention. After the intervention, take a sample to observe whether there is inferior vena cava thrombosis and evaluate the DVT thrombolysis.

[0027] (2) Thrombus length and weight: The condition of DVT is evaluated by measuring the length and weight of the thrombus.

[0028] (3) DVT histology: The lesion status of DVT is evaluated by observing the thrombolysis and fibrosis of the inferior vena cava under a microscope.

[0029] Experimental results are as follows Figure 1-6 As shown.

[0030] Depend on Figure 1 , 2 As shown in sections 3 and 4, the incidence, length, and weight of thrombi in rats treated with pirfenidone were significantly lower than those in the model group after administration, indicating that pirfenidone can promote the dissolution of thrombi in animals.

[0031] Depend on Figure 5 , 6 The results of HE and Masson staining analysis showed that the degree of tissue lesions in the pirfenidone treatment group was significantly lower than that in the model group, indicating that pirfenidone can effectively alleviate DVT in rats with subtotal ligation of the inferior vena cava.

[0032] In summary, pirfenidone has good efficacy against DVT with no adverse reactions, and can alleviate DVT induced by subtotal ligation of the inferior vena cava in rats. It has good application prospects in the treatment, relief or improvement of DVT.

[0033] The above are merely specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. The use of pirfenidone in the preparation of drugs for the treatment of deep vein thrombosis, wherein, The structure of the pirfenidone is shown in formula (Ⅰ): Equation (Ⅰ).

2. The application according to claim 1, characterized in that... The dosage of pirfenidone is 30 mg / kg to 300 mg / kg.

3. The application according to claim 1, characterized in that... Drugs, including pirfenidone, as well as pharmaceutically acceptable salts and excipients of pirfenidone.

4. The application according to claim 3, characterized in that... The pirfenidone salts described are pharmaceutically acceptable salts, including organic and inorganic salts.

5. The application according to claim 4, characterized in that... The organic salts include one or more of methanesulfonates, formates, acetates, trifluoroacetates, maleates, tartrates, succinates, fumarates, citrates, benzenesulfonates, p-methylbenzenesulfonates, naphthalenesulfonates, lactates, and benzoates; the inorganic salts include one or more of hydrochlorides, hydrobroms, sulfates, and phosphates.

6. The application according to claim 5, characterized in that... The dosage form of a drug is selected from tablets, capsules, pills, suppositories, aerosols, granules, powders, injections, syrups, tinctures, elixirs, and films.

7. The application according to claim 5, characterized in that... Drugs can be administered orally, by injection, topically, by spray, or by inhalation.

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