Drug sustained-release preparation for inhibiting vascular intimal hyperplasia as well as preparation method and application of drug sustained-release preparation

Abstract

The invention provides a drug sustained release preparation for inhibiting vascular intimal hyperplasia and a preparation method and application thereof, and belongs to the technical field of biological medicine and vascular surgery. The drug sustained release preparation for inhibiting vascular intimal hyperplasia comprises a temperature-sensitive high polymer material and an MRTF / SRF pathway inhibitor, the temperature-sensitive high polymer material is a Pluronic type triblock copolymer; and the effective component of the MRTF / SRF pathway inhibitor is CCG222740 or CCG203971. The invention further discloses a preparation method of the MRTF / SRF pathway inhibitor. The drug sustained release preparation for inhibiting the vascular intimal hyperplasia can continuously release the MRTF / SRF pathway inhibitor at a fixed point in the AVF and other vascular injury repair process, can reduce at least 50% of neointimal area and vascular intimal-intimal thickness, and effectively inhibits the intimal hyperplasia.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine and vascular surgery, and particularly relates to a drug sustained-release preparation for inhibiting vascular intimal hyperplasia and a preparation method and application thereof. BACKGROUND

[0002] Arteriovenous fistula (AVF) is the first choice of vascular access for hemodialysis patients. However, vascular stenosis and occlusion caused by intimal hyperplasia are the main reasons for long-term loss of function of AVF. Existing measures to inhibit intimal hyperplasia include systemic drug therapy, drug balloon dilation, local injection of drugs, etc., but these methods have defects such as large systemic side effects, short maintenance time of drugs in the local area, and complex drug administration operation.

[0003] Pluronic temperature-sensitive triblock copolymer (such as Pluronic F127) is a fluid at low temperature, and can quickly change into a gel after contacting body temperature, and is suitable for use as a local drug sustained-release carrier. Dissolve or disperse the drug in the liquid Pluronic glue at low temperature, and a stable gel can be formed at the target area under the action of body temperature, continuously releasing the drug to the local tissue, reducing systemic exposure, and prolonging the drug effect time.

[0004] The MRTF / SRF (Myocardin-Related Transcription Factor / Serum Response Factor) signaling pathway regulates the proliferation and migration of vascular smooth muscle cells, and is closely related to intimal hyperplasia. CCG222740 and other small molecule compounds can specifically inhibit the pathway, thereby inhibiting the activation of smooth muscle cells.

[0005] However, there is no report on the local sustained-release application of MRTF / SRF pathway inhibitors through temperature-sensitive polymer materials for preventing intimal hyperplasia after AVF surgery. SUMMARY

[0006] Therefore, the present application aims to provide a drug sustained-release preparation for inhibiting vascular intimal hyperplasia and a preparation method and application thereof.

[0007] In order to achieve the above-mentioned purpose, the present application provides the following technical solutions: One of the technical solutions of the present application is a drug sustained-release preparation for inhibiting vascular intimal hyperplasia, comprising a temperature-sensitive polymer material and a MRTF / SRF pathway inhibitor. The temperature-sensitive polymer material is a Pluronic triblock copolymer. The effective component of the MRTF / SRF pathway inhibitor is CCG222740 or CCG203971.

[0008] In the preferred embodiment of the present application, the drug sustained-release preparation for inhibiting vascular intimal hyperplasia further comprises a buffer solution; the concentration of the MRTF / SRF pathway inhibitor in the drug sustained-release preparation for inhibiting vascular intimal hyperplasia is 200 µM, and the volume ratio of the temperature-sensitive polymer material to the MRTF / SRF pathway inhibitor is 1:50.

[0009] In the preferred embodiment of the present application, the Pluronic triblock copolymer is Pluronic F127 or Pluronic F68, and the MRTF / SRF pathway inhibitor is CCG222740 or CCG203971.

[0010] In the preferred embodiment of the present application, the drug sustained-release preparation for inhibiting vascular intimal hyperplasia is in a liquid state at 0-4℃ and in a gel state at 25-37℃. After forming a gel state, the drug can be released.

[0011] The second technical solution of the present application is a preparation method of the above-mentioned drug sustained-release preparation for inhibiting vascular intimal hyperplasia, comprising the following steps: dissolving the temperature-sensitive polymer material in a buffer solution to obtain solution 1; adding the MRTF / SRF pathway inhibitor to the solution 1 and mixing uniformly to obtain the drug sustained-release preparation for inhibiting vascular intimal hyperplasia.

[0012] In the preferred embodiment of the present application, the temperature of the buffer solution is 0-4℃.

[0013] In the preferred embodiment of the present application, the buffer solution is physiological saline or phosphate buffered physiological saline (PBS).

[0014] In the preferred embodiment of the present application, the mass percentage concentration of the temperature-sensitive polymer material in the solution 1 is 20%-50%.

[0015] In the preferred embodiment of the present application, the mass percentage concentration of the temperature-sensitive polymer material in the solution 1 is 20%, 30%, 40% or 50%.

[0016] After the drug sustained-release preparation for inhibiting vascular intimal hyperplasia is prepared, it is stored at 4℃ for standby use.

[0017] The third technical solution of the present application is the use of the above-mentioned drug sustained-release preparation for inhibiting vascular intimal hyperplasia in the preparation of a drug for treating or preventing vascular stenosis caused by abnormal proliferation of vascular smooth muscle cells.

[0018] In a preferred embodiment of the present application, the vascular stenosis caused by abnormal proliferation of vascular smooth muscle cells includes internal arteriovenous fistula, arterial bypass graft, vascular stent implantation or after angioplasty.

[0019] The drug sustained-release preparation for inhibiting vascular intimal hyperplasia provided by the present application is in liquid state at 0-4 DEG C, and is applied locally or injected into the vascular anastomotic stoma or the predetermined site immediately after the operation, and the local sustained-release preparation forms a gel after contacting with body temperature, and slowly releases the drug for 1-14 days.

[0020] The present application discloses the following technical effects: The drug sustained-release preparation for inhibiting vascular intimal hyperplasia provided by the present application can realize site-specific drug release, inhibit local smooth muscle cell proliferation and migration, and the use method is simple, and can be completed synchronously with the operation.

[0021] The drug sustained-release preparation for inhibiting vascular intimal hyperplasia provided by the present application can realize site-specific, continuous release of MRTF / SRF pathway inhibitors in the process of AVF and other vascular injury repair, can reduce at least 50% of the neointimal area and the vascular media-intima thickness, and effectively inhibit intimal hyperplasia. BRIEF DESCRIPTION OF DRAWINGS

[0022] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed in the embodiments will be briefly introduced below, and obviously, the drawings in the following description can only be some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without any creative effort.

[0023] Figure 1 The model diagram of the drug sustained-release preparation for inhibiting vascular intimal hyperplasia provided by the present application in the application of mouse arteriovenous fistula; Figure 2 The model diagram of the drug sustained-release preparation for inhibiting vascular intimal hyperplasia provided by the present application in the application of human arteriovenous fistula; Figure 3 The left graph in the middle is HE staining of the outflow vein of arteriovenous fistula, and the right graph is the quantification of the vascular media-intima thickness and the neointimal area; Figure 4 The left graph in the middle is a-SMA (ACTA2) immunofluorescence staining of the outflow vein of arteriovenous fistula, and the right graph is the quantification of the a-SMA positive vascular wall thickness. DETAILED DESCRIPTION

[0024] Now, various exemplary embodiments of the present application will be described in detail, and the detailed description should not be considered as a limitation of the present application, but should be understood as a more detailed description of certain aspects, characteristics and embodiments of the present application.

[0025] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the present application. Additionally, for a range of values, the disclosure herein intends each intervening value between the lower and upper limits of that range. Each smaller range that falls within the broader ranges is also contemplated. The upper and lower limits of these smaller ranges can independently be included or excluded in the range, and are also encompassed within the application, subject to any specifically excluded limit in the stated range. Where the described

[0026] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application. All documents mentioned herein are incorporated by reference to disclose and describe in detail the methods and / or materials that are related to the present application. In the case of conflict between the content of the specification and that of any document incorporated herein by reference, the content of the specification controls.

[0027] Many modifications and variations of the present application described in the specification are possible without departing from the scope or spirit of the application. Other embodiments of the application will be apparent to those skilled in the art from consideration of the specification and practice of the application disclosed herein. The specification and examples are illustrative only.

[0028] As used herein, the terms "comprises", "comprising", "includes", "including", "has", "having", "contains", "containing", or variations thereof, are intended to be open-ended terms that mean including, but not limited to.

[0029] The technical solutions described in the present application are conventional solutions in the art, and the reagents or raw materials used are commercially available or have been disclosed, unless otherwise specified.

[0030] In the examples, Pluronic F127 was purchased from Sigma-Aldrich Company, and CCG222740 was purchased from TargetMol Company.

[0031] In order to better understand the present application, the content of the present application is further illustrated below in combination with examples, but the content of the present application is not limited only to the following examples.

[0032] Example 1 Preparation of Pluronic F127-CCG222740 drug sustained-release preparation: Pluronic F127 powder was dissolved in sterile PBS at 4°C and stirred until completely dissolved to obtain a clear liquid with a Pluronic F127 mass fraction of 30%. CCG222740 was then added to the clear liquid and mixed thoroughly to obtain a Pluronic F127-CCG222740 sustained-release formulation (the concentration of CCG222740 in the Pluronic F127-CCG222740 sustained-release formulation was 200 μM). The prepared liquid formulation was dispensed into sterile syringes and stored at 4°C for later use.

[0033] Effect verification: Local administration of the drug after AVF surgery in mice. An abdominal aorta-inferior vena cava (AVF) model was established in C57BL / 6 mice. After surgery, approximately 150 μL of the liquid formulation from Example 1, stored at 4°C, was evenly applied to the anastomosis and the surface of the blood vessel below the renal vein as the experimental group (CCG group). 3 μL of DMSO (dimethyl sulfoxide, a drug cosolvent) was added to 150 μL of Pluronic gel as the control group (Vehicle group). Under body temperature, the liquid formulation rapidly converted to a gel and did not need to be removed postoperatively.

[0034] in conclusion: Samples were taken 14 days post-surgery, and HE and immunofluorescence staining were performed (e.g.) Figure 3 Middle left picture Figure 4 (As shown in the middle left figure) Mice with locally applied Pluronic F127-CCG222740 sustained-release formulation showed significantly lower venous intima thickness than the control group. Pluronic F127 colloid loaded with CCG222740 reduced neointima area and vascular media-intima thickness by at least 50% (e.g., Figure 3 (As shown in the middle right figure).

[0035] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A sustained-release drug formulation for inhibiting vascular intimal hyperplasia, characterized in that, Including thermosensitive polymer materials and MRTF / SRF pathway inhibitors; The temperature-sensitive polymer material is a Pluronic triblock copolymer; The active ingredient of the MRTF / SRF pathway inhibitor is CCG222740 or CCG203971.

2. The sustained-release drug formulation for inhibiting vascular intimal hyperplasia according to claim 1, characterized in that, The sustained-release formulation of the drug for inhibiting vascular intimal hyperplasia also includes a buffer solution; the concentration of the MRTF / SRF pathway inhibitor in the sustained-release formulation of the drug for inhibiting vascular intimal hyperplasia is 200 µM, and the volume ratio of the thermosensitive polymer material to the MRTF / SRF pathway inhibitor is 1:

50.

3. The sustained-release drug formulation for inhibiting vascular intimal hyperplasia according to claim 2, characterized in that, The Pluronic triblock copolymer is Pluronic F127 or Pluronic F68, and the MRTF / SRF pathway inhibitor is CCG222740 or CCG203971.

4. The sustained-release drug formulation for inhibiting vascular intimal hyperplasia according to claim 1, characterized in that, The sustained-release formulation of the drug that inhibits vascular intimal hyperplasia is in liquid form at 0–4°C and in gel form at 25–37°C.

5. A method for preparing a sustained-release drug formulation for inhibiting vascular intimal hyperplasia as described in claim 1, characterized in that, Includes the following steps: The thermosensitive polymer material was dissolved in a buffer solution to obtain solution 1; An MRTF / SRF pathway inhibitor was added to solution 1 and mixed thoroughly to obtain the sustained-release formulation of the drug that inhibits vascular intimal hyperplasia.

6. The preparation method according to claim 5, characterized in that, The temperature of the buffer solution is 0–4°C.

7. The preparation method according to claim 5, characterized in that, The mass percentage concentration of the thermosensitive polymer material in solution 1 is 20%-50%.

8. The use of the sustained-release formulation of the drug for inhibiting vascular intimal hyperplasia as described in claim 1 in the preparation of a drug for treating or preventing vascular stenosis caused by abnormal proliferation of vascular smooth muscle cells.

9. The application according to claim 8, characterized in that, Vascular stenosis caused by abnormal proliferation of vascular smooth muscle cells includes arteriovenous fistulas, arterial bypass grafts, vascular stent implantation, or angioplasty.

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