CN110240542B - Selective β 1-adrenoceptor inhibitor and application thereof in preparation of cardiovascular disease drugs - Google Patents

Abstract

The invention provides a compound with selective β 1-adrenaline inhibition activity, namely, chromane-3, 12E-double bond-15, 16-dicarboxylic acid, of formula (I), wherein the compound has selective β 1-adrenaline receptor inhibition activity, has inhibition activity on β 1 receptor more than six times of that of β 2 receptor, can effectively treat cardiovascular diseases including hypertension, myocardial ischemia, cardiac hypertrophy, coronary heart disease, myocardial infarction, stroke, atherosclerosis, heart failure and angina pectoris, and avoids the side effect of a non-selective β adrenaline receptor inhibitor.

Description

Selective β 1-adrenoceptor inhibitor and application thereof in preparation of cardiovascular disease drugs

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to an β 1-adrenoceptor inhibitor, a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the β 1-adrenoceptor inhibitor.

Background

Cardiovascular disease is an important cause of morbidity and mortality worldwide, and according to the statistics of the World Health Organization (WHO), death due to cardiovascular disease mostly occurs in countries with moderate income, and WHO predicts that cardiovascular disease will be the leading cause of death in developing countries, and therefore, there is an urgent need for treatment and prevention of cardiovascular disease at present, β adrenergic receptor inhibitor is a drug effective for treating cardiovascular disease proved by clinical experiments for many years, β adrenergic receptor can be divided into 3 subtypes (β 1, β, and β 3), β receptor is mainly distributed in heart, accounting for 75-80% of β adrenergic receptor, and related to myocardial contractility, β receptor is mainly distributed in lung, dryness, kidney, peripheral blood vessel, and secretion of epinephrine and relaxation of blood vessel, bronchus, etc. β receptor is mainly distributed in fat tissue, and related to regulation of fatty acid metabolism.

β the main mechanism of use of adrenoceptor inhibitors in the treatment of cardiovascular disease is to block the heart's β 1 adrenoceptors, reducing the force and rate of cardiac contractility and thus the arterial blood pressure and cardiac load in addition, kidney β 1 adrenoceptors block the release of renin, thus reducing angiotensin II and plasma aldosterone levels different β adrenoceptor inhibitors also differ significantly in pharmacodynamic and pharmacokinetic properties.

The first β adrenoceptor inhibitor drug, such as propranolol, is non-specific and blocks both β 1 and β 02 receptors, and light is widely used in hypertensive patients, but experiments have shown that many adverse effects are produced, mainly due to β 2 adrenoceptor being blocked, which adversely affect the respiratory system.the second β 1 adrenoceptor inhibitor, such as atenolol, metoprolol and bisoprolol, has a higher affinity for β 1 receptors, but substantially loses specificity at high doses.metoprolol has no β receptor agonism and little membrane stabilization.bisoprolol, atenolol still has β 1 receptor selectivity at high doses, no intrinsic sympathomimetic activity and membrane stabilization activity.the third generation β 1 adrenoceptor inhibitor, such as labetalol, bucindolol, carvedilol and nebivolol, each has its advantages and disadvantages.therefore there is a need to develop a class of inhibitors with specific blockade at β 1 receptors, with higher β 1 receptor selectivity and low adverse effects.

Hippocampus Laui (Polyalthia Laui) is a kind of arbor belonging to genus Cryptonia (Polyalthia) of Annonaceae family. The plant has over 120 kinds, mainly distributed in tropical and subtropical areas from Thailand, Vietnam, India to Philippine, and there are 17 kinds in China, mainly produced in Taiwan, Hainan, Guangdong and other south areas. Hainan Artocarpus heterophyllus is a special species of Hainan, and has better curative effects on treating diseases such as dysmenorrhea, globus hystericus, qi stagnation and abdominal pain and the like all the time. At present, scholars at home and abroad have obtained diterpenes, triterpenes, annonaceous acetogenins, alkaloids and other chemical components from plants in the genus of the vigna, and have various known medical applications. Wherein the clerodane-3, 12E-double bond-15, 16-dicarboxylic acid shown in the formula (I) is a clerodane type diacid derivative.

The inventor of the application unexpectedly finds that the compound has excellent selective β 1-adrenoceptor inhibition effect in the process of researching an adrenoceptor β receptor inhibitor, and is expected to be used as a drug of β 1-adrenoceptor inhibitor and used for treating cardiovascular diseases, and the compound and pharmaceutically acceptable salts thereof are used as selective β 1-adrenoceptor inhibitor and used as an active compound for treating cardiovascular diseases.

Disclosure of Invention

It is an object of the present invention to provide a selective β 1 adrenergic inhibitor, said β 1 adrenergic inhibitor having selective blocking of the β 1 subtype of epinephrine and having reduced or no inhibitory activity against other subtypes of epinephrine (α, β 2).

The above object of the present invention is achieved by the following technical solutions:

the chemical name of the compound shown in the formula (I) or the pharmaceutically acceptable salt thereof is clerodane-3, 12E-double bond-15, 16-dicarboxylic acid.

The pharmaceutically acceptable salts of the formula (I) are sodium salts, potassium salts, ammonium salts, magnesium salts and calcium salts of the compound of the formula (I), and the pharmaceutically acceptable salts are in the form of single salts, double salts or a combination of the two.

It is a further object of the present invention to provide pharmaceutical compositions of the compounds of formula (I) as pharmaceutically active ingredients.

In one embodiment of the present invention, the agents of the present invention may be used alone or in combination with other agents, such as anti-inflammatory agents, antithrombotic agents, antiplatelet agents, lipid-lowering agents, direct thrombin inhibitors, calcium channel blockers, cyclooxygenase-2 inhibitors and angiotensin system inhibitors.

The pharmaceutically active compounds of the present invention may also be used in the form of pharmaceutically acceptable salts, and suitable addition salts with pharmaceutically acceptable bases include metal, or ammonium salts of calcium, magnesium, potassium, sodium, or salts formed with organic amines such as methylamine, ethylamine, dimethylamine, diethylamine, ethanolamine, N' -dibenzylethylenediamine, choline, meglumine (N-methylglucamine). All of these salts can be prepared from the corresponding compounds by conventional methods, e.g., by reacting the appropriate acid with the compound. The compound of formula (I) provided by the invention has two carboxyl groups, and can be prepared into a single salt, a double salt or a combination of the two according to the actual use requirement. The particular form and salt ratio will be selected by those skilled in the art based on the actual clinical situation.

Salts of the compounds of formula (I) may be prepared by conventional methods known to those skilled in the art, for example by mixing a compound of formula (I) with an inorganic or organic base in a solvent or diluent, or from other salts by cation exchange.

The mode of administration and dosage of the invention are selected according to the particular stage of the disease being treated, the age and condition of the patient, the duration, the particular mode of administration, and the like, as is practical and desired, and within the knowledge and time frame of one skilled in the art. Generally, an effective amount is sufficient to cause a favorable phenotypic change, such as alleviation or elimination of symptoms, permanent cessation of progression, or delay of onset of the disease. In general, the dosage of the active compounds according to the invention is from about 0.01mg/kg to 10mg/kg per day, preferably orally, once or several times a day.

Optionally, the active compounds of the present invention may be formulated into pharmaceutical formulations with a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" refers to one or more compatible solids or liquids, diluents or capsules suitable for administration to the human body. The carrier does not destroy the efficacy of the active compound.

Various modes of administration may be employed so long as they achieve effective levels of the active compound without causing clinically unacceptable side effects. Modes of administration include oral, rectal, topical, intranasal, intradermal, transdermal or parenteral. Oral administration is preferred. Specific modes of administration include tablets, capsules, pills, powders, aqueous or oily solutions or suspensions. Such dosage forms will typically include one or more pharmaceutically active ingredients selected from, for example, adjuvants, carriers, binders, lubricants, diluents, stabilizers, buffers, emulsifiers, surfactants, preservatives, flavoring agents, and coloring agents. The choice of such dosage forms and excipients is well understood by those skilled in the art, and the most appropriate manner of administering the active ingredient will depend on a variety of factors.

Preferably, the pharmaceutical compositions of the present invention are in a form for oral administration. The solid compositions of the present invention are preferably prepared in unit dosage form and are in known pharmaceutical forms such as tablets and capsules. The tablets may suitably be prepared as follows: the effective composition is mixed with any inert diluent in the presence of a disintegrant and a lubricant and the composition is compressed into tablets by known methods. If desired, suitable tablets may be provided, for example enteric coated tablets prepared by known methods. Likewise, capsules containing the effective composition, optionally in the form of granules, may be prepared in a conventional manner and, if desired, enteric-coated capsules may be provided in a known manner. Tablets may be formulated according to methods known in the art to provide controlled release of the compounds of the invention. Other compositions for oral administration also include oily or aqueous suspensions containing a compound of the invention in a vegetable oil.

The compounds or pharmaceutical compositions of the present invention may also be used in the form of injections. The formulation includes an alkaline buffer for adjusting the pH of the aqueous injection formulation between about 7 and about 7.6. Examples of suitable acidic buffers include alkali and base salts such as sodium bicarbonate, potassium bicarbonate, and the like, and alkali metal or ammonium salts of organic acids such as oxalates, maleates, fumarates, lactates, malates, tartrates, citrates, benzoates, acetates, methanesulfonates, tosylates, benzenesulfonates, ethanesulfonates, and the like. The pH of the injection is about 7 to about 7.6, preferably about 7.2 to about 7.4. The active compounds are used in the aqueous injection preparations in amounts of about 0.1 to about 2.5% by weight, preferably about 0.2 to about 1.5% by weight, based on the total weight of the injection preparation.

When the composition is a solid composition, the pharmaceutical composition preferably contains 1-99 wt% of active ingredient, 1-95 wt% of diluent, 0.1-10 wt% of disintegrant, 0.1-10 wt% of lubricant, and optionally 0.1-15 wt% of binder and optionally 0.1-10 wt% of flow promoter.

In a more preferable technical scheme of the invention, the pharmaceutical composition comprises 20-50 wt% of active ingredients, 30-60 wt% of diluents, 1-5 wt% of disintegrants, 1-5 wt% of lubricants, and optionally 2-10 wt% of binders and optionally 1-5 wt% of flow aids. The active ingredient is the compound of the formula (I) and/or pharmaceutically acceptable salt provided by the invention.

As further adjunct ingredients in addition to the active ingredient in the pharmaceutical compositions provided herein, suitable diluents include lactose, tricalcium phosphate, dextrin, sorbitol, mannitol, sucrose, starch (e.g., potato starch, corn starch or amylopectin) mixtures; suitable disintegrants include corn starch, sodium starch glycolate, low-substituted hydroxypropylcellulose, alginic acid, calcium carboxymethylcellulose, croscarmellose sodium or mixtures thereof, preferably corn starch, calcium croscarmellose, sodium carboxymethylcellulose; suitable lubricants include magnesium stearate, calcium stearate, hydrogenated vegetable oils, polyethylene glycols or mixtures thereof, preferably magnesium stearate, stearic acid; suitable binders include polyvinylpyrrolidone, gelatin, hydroxypropylmethylcellulose, starch or mixtures thereof, preferably polyvinylpyrrolidone; suitable glidants include talc, colloidal silicon dioxide or mixtures thereof. It will be appreciated by those skilled in the art that a particular excipient may perform more than one function, for example, corn starch may act as a diluent, binder, or may also act as a disintegrant.

The present invention also relates to the combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof with an antibacterial agent including but not limited to penicillin derivatives, tetracycline, macrolide, β -lactam, fluoroquinolone, an antiviral agent including but not limited to acyclovir, famciclovir, valacyclovir, cidofovir, ribavirin, a cardiovascular agent including but not limited to calcium channel blockers, α -adrenoceptor inhibitors, angiotensin converting enzyme inhibitors, lipid lowering agents.

It is another object of the present invention to provide the use of a compound of formula (I) and/or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of cardiovascular diseases.

β receptor blocker has cardiovascular protection effect, the main mechanism is to resist toxicity of catecholamine adrenergic transmitters, especially cardiac toxicity mediated by β 1 receptors, other mechanisms are also to resist hypertension and myocardial ischemia, play a certain role in blocking renin angiotensin aldosterone system by inhibiting renin release, improve cardiac function and increase left ventricular ejection fraction, resist arrhythmia, etc. β 1 receptor blocker has clinical treatment effect of blood pressure lowering by antagonizing excessive activation of sympathetic nervous system in application of hypertension condition, the main blood pressure lowering mechanism is related to lowering cardiac output, improving blood pressure regulation function of baroreceptors, and inhibiting renin aldosterone system, also prevents cardiotoxicity effect of catecholamine by lowering nervousness, and has clinical treatment application of heart failure by lowering myocardial contractility, heart oxygen consumption and blood pressure, and also plays a role in prolonging angiotensin, prolonging cardiac failure, relieving myocardial ischemia, improving myocardial ischemia, and improving myocardial ischemia.

The compound of formula (I) and the pharmaceutically acceptable salt thereof can be used as β -1 adrenoceptor inhibitor for treating cardiovascular diseases, and have the following advantages:

the compound has excellent β 1-adrenoreceptor inhibition activity, good physicochemical property and stable quality, and is suitable for large-scale industrial production.

Secondly, the compound has selective β 1-adrenoceptor inhibition activity, the inhibition activity to β 1-adrenoceptor is more than six times of that of β 2-adrenoceptor subtype, side effects are avoided, and the safety is high.

Detailed Description

The selective β 1 adrenergic inhibitors of the present invention are further described below in connection with the specific examples, but it should be understood that the scope of the invention is not limited to the following examples.

Unless otherwise defined, all terms of art used hereinafter have the same meaning as commonly understood by one of ordinary skill in the art. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention.

Example 1 the beneficial effects of the active compounds according to the invention are further illustrated below by in vitro pharmacological activity experiments, but this should not be understood as having the following beneficial effects on the compounds according to the invention only

The experimental rats are Wistar rats with unlimited sex and weight of 300 + -25 g, provided by the animal center of the department of medicine of Beijing university.

β 1 receptor membrane protein is prepared from heart tissue of Wistar rat, β 2 receptor membrane protein is prepared from lung tissue of Wistar rat, the preparation method can refer to Willliams, LT method (refer to Willliams, LT: Science,1976,192: 791). The rat is specifically treated by cutting head and bleeding, rat heart and lung tissue are rapidly taken out and put into precooled 0-4 ℃ centrifugal buffer solution, the envelope is stripped, 20 times of centrifugal buffer solution is added, the mixture is crushed by a homogenizer, 10000r/min is crushed for 60s, the mixture is centrifuged for 10 min at 4000G, supernatant is transferred and centrifuged again for 10 min at 1500G, the supernatant is discarded, the precipitate is the receptor-containing membrane protein, the membrane protein is homogenized and suspended by heat preservation buffer solution, and membrane suspension with β 1 receptor or β 2 receptor protein is respectively obtained, and the whole preparation process is carried out at 0-4 ℃ in ice bath.

The test method comprises the following steps: chloramine-T method (see Engel G, Hoyer D) was used.¹²⁵IBE, a new high Affinityradiol and for the adrenocepts, Eur.J.Pharmacol,1985,107(2):111-¹²⁵Separately adding 100 mu L of membrane containing recombinant human epinephrine β receptor, IPIN (about 50pmol/L) and DMSO solutions of compounds of formula (I) in different concentrations into a 96-well plate, adding no compound into a blank control well, incubating at 37 ℃ for 30min, adding 10mL of Tris-HCl (10mmol/L, pH7.4) buffer solution to terminate the reaction, filtering through a glass fiber membrane, washing the membrane once with 10mL of Tris-HCl, draining, and measuring on a gamma counter¹²⁵The scintillation number per minute of I, through Hill plot to find the half effective inhibitory concentration IC50. different concentrations of the compound solution of formula (I) are prepared according to the following method, accurately weighing the compound of formula (I), adding DMSO to dissolve, fully mixing, preparing 10 μ M mother liquor solution, using DMSO to dilute the mother liquor to 5 μ M, 1 μ M, 0.5 μ M, 0.1 μ M, 50M, 10nM, 1nM, 0.5nM, 0.1 nM. step by step, the test results of the inhibitory activity of the compound of formula (I) on the adrenergic receptor β of the invention are shown in Table 351.

TABLE 1

Compound (I)	β1IC50(nM)	β2IC50(nM)
			A compound of formula (I)	203	1264
Atenolol	542	12783

As can be seen from the data in Table 1, the active compound provided by the invention has selective inhibitory activity to β 1-adrenoceptor, has more than six times of inhibitory activity to β 1 receptor than β 2 receptor, has excellent selective β 1-adrenoceptor inhibitory activity, and has good clinical application prospect for treating cardiovascular diseases.

EXAMPLE 2 Effect of Compounds of formula I on the rat model of spontaneous hypertension of SHR

Based on the results of the inhibition of β receptor by the compound of formula I, we further evaluated the hypotensive ability of the compound of formula (I) of the present invention using spontaneous hypertensive rat (SHR rat) model.

The experimental method comprises the following steps: taking 16-week-old SHR rats, and randomly dividing the SHR rats into 4 groups: model group, positive medicine group, administration group, each group is 8. Each group of rats was bred in a conventional environment with free access to water and food, and after 3 days of breeding, administration by gavage was started, atenolol was administered to the positive drug group at 5mg/kg, the compound of formula I was administered at 5mg/kg to the administration group, and an equivalent amount of physiological saline was administered to the model group once a day for 4 weeks.

Blood pressure measurement: the method comprises the steps of detecting blood pressure by a rat tail artery pressure measurement method, detecting by a BP98-A type noninvasive experimental animal blood pressure measuring instrument, specifically, detecting the blood pressure of a tail artery of a rat at about 9 am every day, fixing the rat in a matched rat cage, placing the root of the rat tail into a sleeve of the blood pressure measuring instrument after the rat is quiet, measuring the systolic pressure of the tail artery, continuously measuring for 5 times, wherein the interval of each time is 3 minutes, taking the average value of the systolic pressure of the tail artery as the blood pressure value of the rat, and measuring the rat as the blood pressure basic value 1 day before administration.

The specific results are shown in table 2:

TABLE 2

Or means P <0.05 or 0.01 by T-test compared to model group

As can be seen from Table 2, the compound of formula I provided by the invention has better effect as antihypertensive drug than the known β 1 selective inhibitor atenolol.

EXAMPLE 3 Effect of Compounds of formula I on myocardial ischemia model (LAD) in SD rats

The experimental method comprises the following steps: taking SD rat, about 200g of 8 weeks old; randomized into 4 groups: a sham operation group, a model group, a positive medicine group and a dosing group; each group had 8. The molding method comprises the following steps: pentobarbital sodium is injected into the abdominal cavity for 30mg/kg anesthesia, and the limbs are connected with an electrocardiograph (measuring T2 electrocardiogram) and connected with an animal respirator. The third or fourth intercostal thoracotomy, stripping the pericardium, running the anterior descending branch of the left coronary artery (LAD) with the small round needle 5/0 thread about 2mm below the left auricle, and leading the limb leads or the electrocardiogram of the chest leads to the QRS wave with the ST-segment raised by more than 0.2mV or raised and widened, which indicates the successful ligation of the LAD. The sham group was only threaded and not ligated. After stabilizing for 15 minutes, the chest is sutured, and the rat is separated from the animal respirator to recover spontaneous respiration after suturing. Beginning intragastric administration on the 2 nd day of the model building, and administering atenolol 5mg/kg as a positive medicine group; the administration group is used for administering 5mg/kg of the compound of the formula I; the administration is 1 time daily and is continued for 3 days.

1 day after the administration, the rats are anesthetized, 4ml of blood is taken through an abdominal aorta, the rats are killed after ischemia, the hearts of the rats are taken, after freezing for 15 minutes, the myocardium is taken from the apex to the ligation part in parallel at an angle vertical to the long axis of the heart, one point of the myocardium at the apex of the heart is discarded without use, the rest part is uniformly cut into 5 pieces, the thickness of each piece is about 1mm, and the myocardial infarction area of the rats is measured by adopting a TTC method. Fixed with formaldehyde solution, infarcted myocardium appeared white, non-ischemic myocardium area appeared red. After dyeing IS finished, a digital camera shoots a myocardial section Image, and the ratio of the area (IS) of the infarcted area to the total area IS calculated through Image Pro Plus V6.0 Image processing software, so that the percentage (IS%) of the myocardial infarction area can be calculated.

Centrifuging plasma at 1500rpm, collecting upper layer serum, and measuring LDH and CK-MB content in serum with full-automatic biochemical analyzer

The specific results are as follows:

1. the myocardial infarction areas of the rats are shown in table 3:

TABLE 3

Group of	Artificial operation group	Model set	Positive drug group	Administration set
					IS％	0	42.13±5.24	16.84±4.85**	12.36±5.77**

As can be seen from Table 3, the area of myocardial infarction area of the rats in the administration group is obviously reduced, and the percentage of the area of myocardial infarction is smaller than that of the positive control group, which shows that the compound of the formula I can relieve the symptoms of myocardial ischemia and play a role in protecting the myocardium.

2. Content of LDH and CK-MB in serum

Group of	Artificial operation group	Model set	Positive drug group	Administration set
					LDH(U/L)	1334.6±147.1	2974.2±295.9##	1627.8±185.4**	1504.9±191.3**
CK-MB(U/L)	657.9±32.8	1823.7±214.5##	1169.1±136.7**	1048.4±118.6**

EXAMPLE 4 formulation of pharmaceutical compositions containing active ingredients of Compounds of formula (I)

The pharmaceutical composition of the embodiment comprises the following components in parts by mass:

the composition is prepared by the following steps:

1) sieving and mixing the compound of formula (I) and corn starch to a homogeneous mixture;

2) granulating the mixture with water and drying;

3) mixing the dried granules with sorbitol and calcium stearate to obtain lubricated granules;

4) the lubricated granules are compressed into tablet cores containing 25mg of the compound of formula (I) per tablet core.

5) The tablet cores were coated with a sugar coating containing 2 wt% gum arabic/77 wt% refined sugar/20 wt% calcium sulfate/1 wt% sodium carboxymethyl cellulose.

The above description is only a preferred embodiment of the present invention, and is not intended to limit the embodiments of the present invention, and those skilled in the art can easily make various changes or modifications according to the main concept and spirit of the present invention, and some modifications and alterations of the present invention should fall into the protection scope of the claims of the present invention. Furthermore, although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.

Claims (5)

1. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein the chemical name of the compound of formula (I) is clerodane-3, 12E-di-double bond-15, 16-dicarboxylic acid:

2. use according to claim 1, wherein the pharmaceutically acceptable salt is a metal salt of calcium, magnesium, potassium, sodium, or an ammonium salt.

3. Use according to claim 2, characterized in that the salt is in the form of a mono-salt, a di-salt or a combination of both.

4. Use according to claim 3, characterized in that the salt is the disodium salt of the compound of formula (I).

5. Use according to any one of claims 1 to 4, characterized in that the cardiovascular disease comprises hypertension, myocardial ischemia, cardiac hypertrophy, coronary heart disease, myocardial infarction, stroke, arteriosclerosis, heart failure, angina pectoris.