Pharmaceutical composition for relieving vasospasm and preparation method and application thereof

Abstract

The invention provides a pharmaceutical composition for relieving vasospasm and a preparation method and application of the pharmaceutical composition, the main functional component is an azilsartan-nitroglycerin mixed solution, the concentration of azilsartan is 10 <-6.2 > mol / L to 10 <-3.5 > mol / L, and the concentration of nitroglycerin is 10 <-8 > mol / L to 10 <-4.5 > mol / L; azilsartan is combined with nitroglycerin for use, so that quicker and more continuous diastole can be caused, and the azilsartan has a wide application prospect in the aspect of preparing medicines for relieving vasospasm in coronary heart disease and peripheral vascular disease operations and interventional therapy processes.

Description

Technical Field

[0001] The present invention relates to the fields of coronary heart disease, peripheral vascular intervention and surgery, and in particular to a pharmaceutical composition for relieving vasospasm, its preparation method and application. Background Art

[0002] Coronary heart disease is one of the most common diseases among middle-aged and elderly people. Currently, the treatments for coronary heart disease mainly include: (1) General treatment: alleviating and reducing risk factors, establishing good living habits, immediately resting during the attack period, sedating, supplying oxygen, and promptly sending the patient to the hospital for treatment. (2) Drug treatment: dilating coronary arteries, increasing blood supply, stabilizing atherosclerotic plaques, reducing inflammation, and preventing the formation of thrombus. (3) Percutaneous coronary intervention: This mainly includes percutaneous transluminal coronary angioplasty, coronary artery stenting, coronary atherectomy, laser angioplasty, etc. (4) Surgical treatment: mainly referring to coronary artery bypass grafting. The internal mammary artery is the best graft material and has become the first choice for bypass graft vessels of the left anterior descending branch. Subsequently, some other arterial materials, such as the radial artery, right gastroepiploic artery, inferior epigastric artery, etc., have also been applied to coronary artery bypass grafting.

[0003] During coronary artery bypass grafting, various reasons can cause intraoperative and postoperative graft vasospasm, affecting the postoperative, especially the early surgical effect, and even causing surgical failure. Graft vasospasm is the main cause of patient death. During cardiac surgery clinically, different vasodilators are often used to prevent or reverse vasospasm caused by bypass grafting. Researchers at home and abroad have conducted a large number of studies on anti-vasospasm treatment, and these research works have greatly reduced the occurrence of graft vasospasm. Nevertheless, the phenomenon of graft vasospasm still occurs during or after surgery, and preventing and treating graft vasospasm has become a long-term medical problem to be solved.

[0004] Azilsartan (trade name Azilva), originally developed by Takeda Pharmaceutical Company of Japan, was launched in Japan in 2012. Azilsartan medoxomil (trade name Edarbi, Takeda) is a prodrug that is hydrolyzed to azilsartan after oral administration. In 2011, the US Food and Drug Administration approved azilsartan medoxomil for the treatment of adult hypertension. Azilsartan is the eighth approved ARB drug after candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, and valsartan, and is a preferred drug for patients with hypertension complicated with type II diabetes. Azilsartan competitively and reversibly binds to the AT1 receptor, and its affinity is more than 10,000 times that of AT2, especially for the AT1 receptors in blood vessels, smooth muscles, and adrenal glands.

[0005] Azilsartan exerts its antihypertensive effect by blocking the binding of Ang II to the AT1 receptor. Different clinical trial data have confirmed that Azilsartan has a stronger antihypertensive effect on systolic blood pressure than Valsartan, Olmesartan, and Ramipril. Azilsartan is more effective than Candesartan in reducing ambulatory systolic and diastolic blood pressure and can continuously lower blood pressure for 24 hours. Azilsartan can exert an insulin resistance effect and prevent cardiovascular remodeling and dysfunction in diabetic patients. For patients with type 2 diabetes complicated with hypertension, Azilsartan can reduce the cardiovascular risk of patients and also improve glucose metabolism and lower fasting blood glucose. And clinical trials have confirmed that healthy people have good safety and tolerance to Azilsartan ester. Although Azilsartan has been on the market, so far, it has not been developed into a solution for relieving vasospasm during the surgical treatment of coronary heart disease and peripheral vascular diseases. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to provide a pharmaceutical composition for relieving vasospasm.

[0007] Another technical problem to be solved by the present invention is to provide a preparation method of the above-mentioned pharmaceutical composition for relieving vasospasm.

[0008] Another technical problem to be solved by the present invention is to provide the application of the above-mentioned pharmaceutical composition for relieving vasospasm.

[0009] To solve the above technical problems, the technical solution of the present invention is:

[0010] A pharmaceutical composition for relieving vasospasm, named AG solution, the main active ingredient is a mixed solution of Azilsartan - Glyceryl trinitrate (also known as Nitroglycerin), wherein the concentration of Azilsartan is 10 -6.2 mol / L - 10 -3.5 mol / L, and the concentration of Glyceryl trinitrate is 10 -8 mol / L - 10 -4.5 mol / L.

[0011] Preferably, for the above-mentioned pharmaceutical composition for relieving vasospasm, the concentration of Azilsartan is 10 -3.5 mol / L, and the concentration of Glyceryl trinitrate is 10 -4.5 mol / L.

[0012] Preferably, for the above-mentioned pharmaceutical composition for relieving vasospasm, it is composed of Ringer's solution, Azilsartan, Glyceryl trinitrate injection, Heparin sodium injection, and Sodium bicarbonate. Among them, the final concentration of Azilsartan in every 500 ml of Ringer's solution is 10 - 6.2 mol / L - 10 -3.5mol / L; the final concentration of nitroglycerin injection is 10 -8 mol / L - 10 -4.5 mol / L; the final concentration of heparin sodium injection is 1700 U / L; the final concentration of sodium bicarbonate is 1 g / L.

[0013] Preferably, in the above drug composition for relieving vasospasm, the final concentration of azilsartan in every 500 ml of Ringer's solution is 10 -3.5 mol / L; the final concentration of nitroglycerin injection is 10 -4.5 mol / L; the final concentration of heparin sodium injection is 1700 U / L; the final concentration of sodium bicarbonate is 1 g / L.

[0014] In the above drug composition, Ringer's solution is a solution, and azilsartan and nitroglycerin are the main solutes. The azilsartan has the molecular formula C 25 H 20 N 4 O 5 , and the molecular weight is 456.45; the nitroglycerin injection has the molecular formula C 3 H 5 N 3 O 9 ; the heparin sodium solution has the molecular formula ca.(C 12 H 16 NS 2 Na 3 ) 20 ; the sodium bicarbonate has the molecular formula NaHC0 3 , and the molecular weight is 84.01.

[0015] Preferably, in the above drug composition for relieving vasospasm, in every 150 ml of the Ringer's solution: 20.8 mg of azilsartan, 6.5 mg of nitroglycerin, 250 U of heparin sodium, 0.5 ml of 8.4% sodium bicarbonate, and the pH value is 7.4.

[0016] The preparation method of the above drug composition for relieving vasospasm specifically comprises the following steps (taking the preparation of 150 ml as an example):

[0017] (1) Weigh 150 ml of Ringer's solution;

[0018] (2) Weigh 20.8 mg of azilsartan, 8.6 μl of nitroglycerin injection, and 40 μl of heparin sodium injection and add them into 150 ml of Ringer's solution respectively;

[0019] (3) Weigh 0.84 g of sodium bicarbonate, add 10 ml of distilled water to obtain an 8.4% solution, and take 0.5 ml and add it into 150 ml of Ringer's solution.

[0020] Use of the above-mentioned pharmaceutical composition in the preparation of a medicament for relieving vasospasm.

[0021] Preferably, in the use of the above-mentioned pharmaceutical composition, azilsartan and nitroglycerin act synergistically, and the medicament prepared for relieving vasospasm is a medicament for relieving vasospasm during the surgery or interventional treatment of coronary heart disease and peripheral vascular diseases.

[0022] Preferably, in the use of the above-mentioned pharmaceutical composition, the surgeries for coronary heart disease and peripheral vascular diseases include coronary artery bypass grafting, percutaneous coronary intervention, and other peripheral vascular interventions, etc.

[0023] The beneficial effects of the present invention are:

[0024] The above-mentioned pharmaceutical composition (AG solution) for relieving vasospasm was studied for its pharmacological effects on the graft vessels used in the surgeries of coronary heart disease and peripheral vascular diseases. It was found that the synergistic effect of azilsartan and nitroglycerin at a specific concentration can more rapidly relieve vasospasm and dilate blood vessels, and increase the final degree of dilation. Vasospasm of the graft vessels after coronary artery bypass grafting has always been a problem. The AG solution at a specific concentration not only has direct significance in the clinical prevention and treatment of anti-vasospasm, but also has a good effect on the medium- and long-term patency rate of the blood vessels after surgery. At the same time, it can also lay a scientific foundation for the study of the mechanism of anti-vasospasm.

[0025] The azilsartan contained in the composition solution for relieving vasospasm obtained through the above research has the effects of lowering blood pressure, protecting the kidneys, reducing inflammatory reactions, and reducing endothelial damage; nitroglycerin is an exogenous nitric oxide donor and a vasodilator drug. The combination of azilsartan and nitroglycerin acts synergistically, and the use can cause a more rapid and sustained dilation. Description of the Drawings

[0026] Figure 1 It is the vasodilation curve when potassium chloride is a vasoconstrictor;

[0027] Figure 2 It is the vasodilation curve when U46619 is a vasoconstrictor;

[0028] Figure 3 It is the antagonistic effect of azilsartan combined with nitroglycerin pre-incubated with blood vessels on potassium chloride;

[0029] Figure 4 It is the antagonistic effect of azilsartan combined with nitroglycerin pre-incubated with blood vessels on U46619. Detailed Embodiments

[0030] To enable those skilled in the art to better understand the technical solution of the present invention, the following further details the technical solution of the present invention in conjunction with the accompanying drawings and specific embodiments.

[0031] In the following drug composition, Ringer's solution is the main solvent, purchased from Otsuka Pharmaceutical Co., Ltd., Guangdong, with a specification of 500 ml; nitroglycerin injection is purchased from Shanxi Kangbao Bioproducts Co., Ltd., with a specification of 1 ml: 5 mg; heparin sodium injection is purchased from Jiangsu Wanbang Biochemical Pharmaceutical Group Co., Ltd., with a specification of 2 ml: 12,500 units.

[0032] Example 1

[0033] A drug composition (AG solution) for relieving vasospasm, composed of Ringer's solution (main solvent), azilsartan, nitroglycerin, heparin sodium, and sodium bicarbonate, wherein:

[0034] Ringer's solution: 150 ml

[0035] Azilsartan: 20.8 mg (final concentration is 10 -3.5 mol / L)

[0036] Nitroglycerin: 6.5 mg (final concentration is 10 -4.5 mol / L)

[0037] Heparin sodium: 250 U (final concentration is 1700 U / L)

[0038] 8.4% sodium bicarbonate: 0.5 ml (final concentration is 1 g / L)

[0039] pH value: 7.4.

[0040] The Ringer's solution has a specification of 500 ml; the azilsartan has a molecular formula of C 25 H 20 N 4 O 5 and a molecular weight of 456.45; the nitroglycerin injection has a specification of 1 ml: 5 mg and a molecular formula of C 3 H 5 N 3 O 9 ; the heparin sodium solution has a specification of 2 ml: 12,500 units and a molecular formula of ca.(C 12 H 16 NS 2 Na 3 ) 20 and a specification of 12,500 units; the sodium bicarbonate has a molecular formula of NaHCO 3 and a molecular weight of 84.01.

[0041] The preparation method of the above-mentioned drug composition (AG solution) for relieving vasospasm is as follows:

[0042] (1) Weigh 150 ml of Ringer's solution;

[0043] (2) Weigh 20.8 mg of azilsartan, 8.6 μl of nitroglycerin injection, and 40 μl of heparin sodium injection and add them to 150 ml of Ringer's solution respectively;

[0044] (3) Weigh 0.84 g of sodium bicarbonate, add 10 ml of distilled water to obtain an 8.4% solution, and take 0.5 ml and add it to 150 ml of Ringer's solution.

[0045] Example 2

[0046] (1) Collection and pretreatment of the internal mammary artery:

[0047] Put the distal end of the graft vessel (internal mammary artery) discarded during coronary artery bypass grafting into the Krebs buffer solution at 4°C filled with a 95% CO 2 , 5% O 2 mixed gas, and transfer it to the laboratory as soon as possible. Carefully separate the peripheral tissues in the buffer solution with dissecting scissors to expose the blood vessel, segment the blood vessel, and each segment is 3 mm. Hang each segment of the blood vessel loop in the bath of a microvascular tension measuring instrument (Model 620, DMT Company, Denmark) and balance for at least 45 minutes.

[0048] (2) Vascular functional experiment:

[0049] Restoration of vascular state:

[0050] Add 6 ml of Krebs buffer solution to the bath of the microvascular tension measuring instrument, set the temperature at 37°C, and continuously introduce 95% CO 2 , 5% O 2 mixed gas until the end of the experiment. Put the separated segment of the blood vessel on the hanging needle used to measure the vascular tension in the bath to make the blood vessel appropriately tense and maintain this state for 1 - 1.5 hours to restore the vascular tension unchanged caused during the transfer and separation process.

[0051] Establishment of the vascular Resting Force state:

[0052] After the blood vessels have had sufficient rest, turn the micrometer to dilate the blood vessels. The dilation process is carried out step by step. Generally, the blood vessel wall pressure reaches 100 mmHg (13.3 kPa) through 3 - 4 dilations. At this time, the inner diameter of the blood vessel L100 is calculated by software simulation. Adjust the micrometer to 0.9 * L100, relax the blood vessel. When the blood vessel is in equilibrium, the Resting Force state of the blood vessel is established. At this time, the dilation state of the blood vessel in vitro is closest to that in vivo. Record the transmural pressure and the inner diameter D100 in this state. After the blood vessel reaches the Resting Force state, to restore the change in tension caused during the stretching process of the blood vessel, this state should be maintained for 1 h to fully restore the blood vessel function.

[0053] To study the influence of non - treatable factors on the vasodilatory function of blood vessels, the experiment was divided into 2 parts:

[0054] Part 1: Relaxation of the internal mammary artery by the AG solution after contraction with potassium chloride

[0055] Cut the same internal mammary artery into 3 segments. After pre - contracting the internal mammary artery with potassium chloride (30 mM, n = 8), add 10 -3.5 mol / L azilsartan, 10 -4.5 mol / L nitroglycerin, and the AG solution described in Example 1. Compare the characteristics of the relaxation of the internal mammary artery by the three groups of solutions. The results are shown in Figure 1 , and the maximum relaxation degrees are 72.7%, 77.3%, and 84.2% respectively. After relaxation with the AG solution, the relaxation speed is fast and persistent.

[0056] Part 2: Relaxation of the internal mammary artery by the AG solution after contraction with U46619

[0057] Cut the same internal mammary artery into 3 segments. After pre - contracting the internal mammary artery with U46619 (20 nM, n = 8), add 10 -3.5 mol / L azilsartan, 10 -4.5 mol / L nitroglycerin, and the AG solution described in Example 1. Compare the characteristics of the relaxation of the internal mammary artery by the three groups of solutions. The results are shown in Figure 2 , and the maximum relaxation degrees are 94.3%, 80.9%, and 98.8% respectively. After relaxation with the AG solution, the relaxation speed is fast and persistent.

[0058] Example 3

[0059] A pharmaceutical composition (AG solution) for relieving vasospasm, which is the same as in Example 1, except that: the concentration of azilsartan is 10 -6.2 mol / L and the concentration of nitroglycerin is 10 -8 mol / L.

[0060] Example 4

[0061] Antagonistic effect of AG solution on potassium chloride-induced vasoconstriction

[0062] A segment of internal mammary artery was cut into 4 pieces and incubated in distilled water (control group), 10 -6.2 mol / L azilsartan, 10 -8 mol / L nitroglycerin, and AG solution (10 -6.2 mol / L azilsartan + 10 -8 mol / L nitroglycerin) for 1 hour. Then, the antagonistic effects of the four groups of blood vessels on potassium chloride-induced vasoconstriction with gradient concentrations (5 mM - 120 mM, n = 6) were compared.

[0063] The results are shown in Figure 3 , and compared with the other three groups ( Figure 3 A), the AG group significantly inhibited the vasoconstrictive effect of potassium chloride (p < 0.001, compared with the control group; p < 0.001, compared with nitroglycerin alone; p < 0.001, compared with azilsartan alone). At 120 mM potassium chloride, there were also significant differences in the maximum contraction value (MAX contraction) of blood vessels ( Figure 3 B, p < 0.01, compared with the control group; p < 0.01, compared with nitroglycerin alone). The concentration for half-maximal effect (EC 50 ) also increased significantly ( Figure 3 C, p < 0.05, compared with the control group).

[0064] Example 5

[0065] Antagonistic effect of AG solution on U46619-induced vasoconstriction

[0066] A segment of internal mammary artery was cut into 4 pieces and incubated in distilled water (control group), 10 -6.2 mol / L azilsartan, 10 -8 mol / L nitroglycerin, and AG solution (10 -6.2 mol / L azilsartan + 10 -8 mol / L nitroglycerin) for 1 hour. Then, the antagonistic effects of the four groups of blood vessels on U46619-induced vasoconstriction with gradient concentrations (10 -5.5 mol / L - 10 -9 mol / L, n = 6) were compared.

[0067] The results are shown in Figure 4 , and compared with the other three groups ( Figure 4(A) significantly inhibited the contractile effect of potassium chloride (p < 0.001, compared with the control group; p < 0.001, compared with nitroglycerin alone; p < 0.001, compared with azilsartan alone). At U4661910 -5.5 mol / L, there were also significant differences in the maximum contraction value (MAX aontraction) of blood vessels ( Figure 4 B, p < 0.001, compared with the control group; p < 0.05, compared with nitroglycerin alone). The median effective concentration (EC 50 ) also increased significantly ( Figure 4 C, p < 0.05, compared with the control group; p < 0.01, compared with nitroglycerin alone; p < 0.05, compared with azilsartan alone).

[0068] The above detailed description of the pharmaceutical composition for relieving vasospasm, its preparation method and application with reference to specific embodiments (including the concentration of each drug in the drug combination) is illustrative rather than restrictive. Several embodiments can be listed within the defined scope. Therefore, changes and modifications without departing from the general concept of the present invention shall fall within the protection scope of the present invention.

Claims

1. A pharmaceutical composition for relieving vasospasm, characterized in that: The main active ingredient is azilsartan-nitroglycerin mixed solution, in which the concentration of azilsartan is 10 -6.2 mol / L-10 -3.5 mol / L, nitroglycerin concentration is 10 -8 mol / L-10 -4.5 mol / L.

2. The pharmaceutical composition for relieving vasospasm according to claim 1, characterized in that: The concentration of azilsartan is 10 -3.5 mol / L, nitroglycerin concentration is 10 -4.5 mol / L.

3. The pharmaceutical composition for relieving vasospasm according to claim 1, characterized in that: It is composed of Ringer's solution, azilsartan, nitroglycerin injection, heparin sodium injection and sodium bicarbonate, wherein the final concentration of azilsartan in each 500ml of Ringer's solution is 10 -6.2 mol / L-10 -3.5 mol / L; the final concentration of nitroglycerin injection is 10 -8 mol / L-10 -4.5 mol / L; the final concentration of heparin sodium injection is 1700U / L; the final concentration of sodium bicarbonate is 1g / L.

4. The pharmaceutical composition for relieving vasospasm according to claim 3, characterized in that: The final concentration of azilsartan in 500 ml of Ringer's solution is 10 -3.5 mol / L; the final concentration of nitroglycerin injection is 10 -4.5 mol / L; the final concentration of heparin sodium injection is 1700U / L; the final concentration of sodium bicarbonate is 1g / L.

5. The pharmaceutical composition for relieving vasospasm according to claim 3 or 4, characterized in that: Every 150 ml of the Ringer's solution contains: 20.8 mg of azilsartan, 6.5 mg of nitroglycerin, 250 U of sodium heparin, 0.5 ml of 8.4% sodium bicarbonate, and a pH value of 7.

4.

6. A method for preparing the pharmaceutical composition for relieving vasospasm according to any one of claims 1 to 5, characterized in that: The specific steps are: (1) Weigh Ringer's solution; (2) Weigh azilsartan, nitroglycerin injection, and heparin sodium injection according to the amount of each group and add them into Ringer's solution; (3) Weigh sodium bicarbonate and add it to distilled water to make a sodium bicarbonate solution, then add it to Ringer’s solution.

7. Use of the pharmaceutical composition according to any one of claims 1 to 5 in the preparation of a drug for relieving vasospasm.

8. The use of the pharmaceutical composition according to claim 7, characterized in that: The prepared medicine for relieving vascular spasm is a medicine for relieving vascular spasm during surgery or interventional treatment of coronary heart disease and peripheral vascular disease.

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