Pharmaceutical composition containing spironolactone and amygdalin for promoting blood circulation, relieving swelling and easing pain and application of pharmaceutical composition

Abstract

The invention belongs to the technical field of medicines, and provides a blood-activating, swelling-eliminating and pain-relieving pharmaceutical composition containing spirolactone and amygdalin and application of the blood-activating, swelling-eliminating and pain-relieving pharmaceutical composition. The pharmaceutical composition is prepared from spirolactone and amygdalin as raw materials according to a certain weight percentage. Experimental results show that the pharmaceutical composition disclosed by the invention has a remarkable synergistic effect in the aspect of pain threshold of mice painful by a hot plate, in the aspect of mouse ear swelling degree caused by xylene and in the aspect of mouse abdominal cavity capillary permeability caused by acetic acid, has no toxic or side effect and has a relatively good clinical application prospect.

Description

Technical Field

[0001] The invention belongs to the technical field of medicines, and in particular relates to a blood circulation promoting, swelling reducing and analgesic medicinal composition comprising spironolactone and amygdalin and an application thereof. Background Art

[0002] Spironolactone is an aldosterone receptor antagonist that can compete with aldosterone for binding to the mineralocorticoid receptor, making it difficult for the aldosterone-receptor complex to form, and can be used to inhibit the pathological effects of aldosterone. Clinically, studies have shown that spironolactone can be used to treat hypertension, chronic kidney disease, diabetes and other diseases.

[0003] Amygdalin is mainly found in bitter almonds, and its extraction methods are diverse, including water extraction, alcohol extraction, macroporous adsorption resin separation and purification, ultrasonic method and microwave method. Amygdalin is now used as an auxiliary drug for relieving cough and asthma, improving immunity and anti-tumor.

[0004] Soft tissue injury refers to a large category of trauma syndromes caused by direct or indirect violence to soft tissue or skeletal muscles, or long-term chronic strain. After the tissue is injured, microcirculation disorders and aseptic inflammation occur, resulting in local swelling and pain. Traumatic injuries and tendon injuries and fractures refer to soft tissue injuries or fractures of the human body caused by external force or excessive force, with swelling and pain as the main symptoms. Traumatic injuries include knives, guns, beatings, sprains, stab wounds, abrasions, sports injuries, etc. The injured parts are mostly painful, swollen, bleeding or fractures, dislocations, etc., and also include some internal organ injuries. Here, soft tissue injuries are the main focus. Western medicine usually uses antibiotics and anti-inflammatory, swelling and analgesic, debridement and suturing, traction, and open reduction in the treatment of various soft tissue injuries, traumatic injuries, and tendon injuries and fractures.

[0005] There is no related report in the prior art that a pharmaceutical composition comprising spironolactone and amygdalin as claimed in the present invention is used for promoting blood circulation, reducing swelling and relieving pain. Summary of the invention

[0006] The purpose of the present invention is to solve the deficiencies of the prior art, to develop and utilize drugs using modern drug research methods, and to combine a large number of pharmacodynamic experimental screenings to provide a drug composition for promoting blood circulation, reducing swelling and relieving pain with synergistic effects, thereby providing a new drug composition for promoting blood circulation, reducing swelling and relieving pain in clinical practice.

[0007] In a first aspect, the present invention provides a pharmaceutical composition for promoting blood circulation, reducing swelling and relieving pain comprising spironolactone and amygdalin, wherein the pharmaceutical composition is composed of spironolactone and amygdalin, and the weight ratio of spironolactone to amygdalin is 1:8-12.

[0008] Furthermore, the weight ratio of spironolactone to amygdalin in the pharmaceutical composition is 1:10.

[0009] In a second aspect, the present invention provides a method for preparing the pharmaceutical composition for promoting blood circulation, reducing swelling and relieving pain as described in the first aspect, comprising mixing the two active ingredients as described in the first aspect.

[0010] In a third aspect, the present invention provides a pharmaceutical preparation for promoting blood circulation, reducing swelling and relieving pain, wherein the pharmaceutical preparation is prepared by mixing the pharmaceutical composition described in the first aspect with pharmaceutically acceptable excipients.

[0011] Furthermore, the auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, retention agent, lubricant, dispersant, plasticizer, opacifier and antioxidant.

[0012] Furthermore, the pharmaceutical preparation is an oral preparation or an injection.

[0013] Furthermore, the oral preparation is a powder, tablet, pill, capsule, granule or oral liquid.

[0014] In the fourth aspect, the present invention provides the use of the blood-activating, stasis-removing, swelling-reducing and analgesic pharmaceutical composition of the first aspect or the blood-activating, stasis-removing, swelling-reducing and analgesic pharmaceutical preparation of the third aspect in the preparation of blood-activating, stasis-removing, swelling-reducing and analgesic drugs.

[0015] The present invention also provides the use of the blood circulation promoting, swelling reducing and analgesic pharmaceutical composition described in the first aspect or the blood circulation promoting, swelling reducing and analgesic pharmaceutical preparation described in the third aspect in the preparation of drugs for treating traumatic injuries, tendon injuries and fractures, and soft tissue injuries.

[0016] The pharmaceutical composition of the present invention effectively improves the high viscosity, high polymerization and high concentration state of blood during acute soft tissue injury and blood stasis, so that the blood stasis state is relieved, thereby achieving the function of promoting blood circulation and removing blood stasis.

[0017] The pharmaceutical composition of the present invention reduces the expression of PGE2 in the serum of rats with acute soft tissue injury, alleviates inflammatory symptoms such as swelling, fever, and pain in local tissues, and helps to recover from acute soft tissue injury; the pharmaceutical composition of the present invention reduces the expression of IL-6 in the serum of rats with acute soft tissue injury, reduces the inflammatory response, and accelerates the repair process of muscle tissue.

[0018] Compared with the prior art, the present invention has the following advantages:

[0019] The present invention discovers for the first time that the pharmaceutical composition composed of spironolactone and amygdalin has a significant synergistic effect in promoting blood circulation, reducing swelling and relieving pain, and provides a new pharmaceutical composition for promoting blood circulation, reducing swelling and relieving pain.

[0020] Pharmacological experiments have shown that the pharmaceutical composition of the present invention exhibits significant synergistic effects in terms of the pain threshold of mice induced by hot plate pain, the swelling of mice ears caused by xylene, and the permeability of mouse peritoneal capillaries caused by acetic acid, and has no toxic side effects, and has good clinical application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The effect of the pharmaceutical composition of the present invention on the pain induced by hot plate in mice was compared with the model group. * P<0.05, ** P<0.01; compared with the combination group, # P<0.05.

[0022] Figure 2 The effect of the pharmaceutical composition of the present invention on ear swelling in mice was compared with the model group. * P<0.05, ** P<0.01; compared with the combination group, # P<0.05.

[0023] Figure 3 The effect of the pharmaceutical composition of the present invention on the permeability of peritoneal capillaries in mice was compared with that of the model group. * P<0.05, ** P<0.01; compared with the combination group, # P<0.05.

[0024] Figure 4 The effect of the pharmaceutical composition of the present invention on the whole blood viscosity of the rat acute soft tissue injury model was compared with the model group. * P<0.05, ** P<0.01; compared with the combination group, # P<0.05.

[0025] Figure 5 The effect of the pharmaceutical composition of the present invention on the plasma viscosity of the rat acute soft tissue injury model was compared with the model group. * P<0.05, ** P<0.01; compared with the combination group, # P<0.05.

[0026] Figure 6 The effect of the pharmaceutical composition of the present invention on serum PEG2 in rats with acute soft tissue injury model was compared with the model group. * P<0.05, ** P<0.01; compared with the combination group, # P<0.05.

[0027] Figure 7The effect of the pharmaceutical composition of the present invention on serum IL-6 in rats with acute soft tissue injury model was compared with the model group. * P<0.05, ** P<0.01; compared with the combination group, # P<0.05. DETAILED DESCRIPTION

[0028] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention.

[0029] Unless otherwise indicated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by those of ordinary skill in the field of the invention or the field in which the terms are used. Although any methods, conditions, substances or materials similar to or equivalent to those disclosed herein may be used in the practice of the present invention, preferred methods, conditions, substances or materials are described herein.

[0030] Example 1 Analgesic effect of the pharmaceutical composition of the present invention on hot plate-induced pain in mice

[0031] 1. Experimental Methods

[0032] Female Kunming mice of clean grade, weighing 18-20 g, were given free access to water and food, and the room temperature was maintained at 20-25°C.

[0033] The temperature of the constant temperature water bath was set to 55°C. When the water temperature rose to (55±0.5)°C, a 500mL beaker was placed in the water bath and preheated for 10 minutes. One mouse was taken each time and placed in the beaker. The time required for its feet to touch the bottom of the beaker and lick its hind feet was recorded as the pain threshold of the mouse. Those who did not lick their hind feet within 30 seconds were discarded. After the mice were marked, the basal pain threshold was measured, and the measurement was repeated twice, and the average was taken as the basal pain threshold. 32 qualified mice were randomly divided into spironolactone group, amygdalin group, combination group, and model group, with 8 mice in each group. Each group of mice was gavaged: the spironolactone group was gavaged with 1.0 mg / kg spironolactone each time, the amygdalin group was gavaged with 10.0 mg / kg amygdalin each time, the combination group was gavaged with 0.5 mg / kg spironolactone + 5.0 mg / kg amygdalin each time, and the model group was gavaged with 0.2 mL of normal saline each time. The drugs were administered continuously for 3 days, 3 times a day, and the pain threshold was measured 90 minutes after the last dose.

[0034] 2. Statistical methods

[0035] Graphpad Prism 9.0 software was used for data processing, and the measured data were expressed as mean ± standard deviation. One-way analysis of variance was used for comparison among multiple groups, and SNK-q was used for pairwise comparison among groups. The difference was considered statistically significant when P < 0.05.

[0036] 3. Experimental results

[0037] Effects on hot plate-induced pain in mice

[0038] Compared with the model group, the combination group significantly increased the pain threshold of the mice induced by hot plate pain, and its effect of increasing the pain threshold of the mice induced by hot plate pain was significantly better than that of the spironolactone alone group and the amygdalin alone group.

[0039] The pharmaceutical composition of the present invention can significantly increase the pain threshold of mice induced by hot plate pain. The two active ingredients, spironolactone and amygdalin, show significant synergistic effects in increasing the pain threshold. Figure 1 .

[0040] Example 2 Effect of the pharmaceutical composition of the present invention on ear swelling in mice

[0041] 1. Experimental Methods

[0042] Thirty-two male mice of Kunming clean grade, weighing 18-20 g, were given free access to water and food, and the room temperature was maintained at 20-25°C.

[0043] Mice were randomly divided into 4 groups: spironolactone group, amygdalin group, combination group, and model group, with 8 mice in each group. Each group of mice was gavaged, with 1.0 mg / kg spironolactone per mouse in the spironolactone group, 10.0 mg / kg amygdalin per mouse in the amygdalin group, 0.5 mg / kg spironolactone + 5.0 mg / kg amygdalin per mouse in the combination group, and 0.2 mL normal saline per mouse in the model group. The drugs were administered for 7 consecutive days, once a day.

[0044] 1 hour after the last administration, 0.03 mL of xylene was applied to the right auricle of each mouse. The mice were killed 0.5 hours later, and the left and right auricles were punched out with a stainless steel punch with a diameter of 8 mm, weighed, and the swelling rate was calculated. Ear swelling rate (%) = (right ear weight - left ear weight) / left ear weight × 100%.

[0045] 2. Statistical methods

[0046] Graphpad Prism 9.0 software was used for data processing, and the measured data were expressed as mean ± standard deviation. One-way analysis of variance was used for comparison among multiple groups, and SNK-q was used for pairwise comparison among groups. The difference was considered statistically significant when P < 0.05.

[0047] 3. Experimental results

[0048] Effects on ear swelling in mice

[0049] Compared with the model group, the combination group significantly inhibited the ear swelling of mice caused by xylene, and its effect of inhibiting the ear swelling of mice caused by xylene was significantly better than that of the spironolactone alone group and the amygdalin alone group.

[0050] The pharmaceutical composition of the present invention can significantly inhibit the ear swelling of mice caused by xylene. The two active ingredients, spironolactone and amygdalin, show significant synergistic effects in inhibiting the ear swelling of mice caused by xylene. Figure 2 .

[0051] Example 3 Effect of the pharmaceutical composition of the present invention on the permeability of peritoneal capillaries in mice

[0052] 1. Experimental Methods

[0053] 32 Kunming clean-grade female mice, weighing 18-20 g, were given free access to water and food, and the room temperature was maintained at 20-25°C.

[0054] Mice were randomly divided into 4 groups: spironolactone group, amygdalin group, combination group, and model group, with 8 mice in each group. Each group of mice was gavaged, with 1.0 mg / kg spironolactone per mouse in the spironolactone group, 10.0 mg / kg amygdalin per mouse in the amygdalin group, 0.5 mg / kg spironolactone + 5.0 mg / kg amygdalin per mouse in the combination group, and 0.2 mL normal saline per mouse in the model group for 3 consecutive days, twice a day. 30 minutes after oral administration on the third day, the mice were fixed with a homemade mouse fixer to expose their tails, and the tails were wiped with 75% alcohol. After the tail veins of the mice were obviously filled, 0.5% Evans blue solution (0.1 mL / 10 g) was injected into the tail vein, and 0.6% acetic acid solution (0.2 mL per mouse) was injected intraperitoneally to induce inflammation. After 20 minutes, the mice were killed by decapitation and dislocation, the peritoneum was cut open, and the abdominal cavity was flushed several times with 5 mL of distilled water. 5 mL of the abdominal flushing fluid was taken with a pipette, and the solution was centrifuged for 5 minutes (3000 r / min). The absorbance (A value) was measured at 590 nm with a UV-visible photometer.

[0055] 2. Statistical methods

[0056] Graphpad Prism 9.0 software was used for data processing, and the measured data were expressed as mean ± standard deviation. One-way analysis of variance was used for comparison among multiple groups, and SNK-q was used for pairwise comparison among groups. The difference was considered statistically significant when P < 0.05.

[0057] 3. Experimental results

[0058] Effects on peritoneal capillary permeability in mice

[0059] Compared with the model group, the combination group significantly inhibited the increase in vascular permeability caused by inflammatory factors, and its effect of inhibiting the increase in vascular permeability caused by inflammatory factors was significantly better than the spironolactone alone group and the amygdalin alone group.

[0060] The pharmaceutical composition of the present invention can significantly inhibit the increase in vascular permeability caused by inflammatory factors. The two active ingredients, spironolactone and amygdalin, show significant synergistic effects in inhibiting the increase in vascular permeability caused by inflammatory factors. Figure 3 .

[0061] The anti-inflammatory and analgesic effects of the pharmaceutical composition of the present invention were studied by the effects of the pharmaceutical composition of the present invention on the hot plate test of mice, the ear swelling test of mice and the permeability of the peritoneal capillaries of mice.

[0062] The pharmaceutical composition group of the present invention can significantly increase the pain threshold of mice induced by hot plate pain, inhibit the ear swelling of mice caused by xylene, and has a significant inhibitory effect on the increase in the permeability of the mouse abdominal capillaries caused by acetic acid.

[0063] The pharmaceutical composition group of the present invention has stronger anti-inflammatory, analgesic and microcirculation improving protective effects than the spironolactone single-use group and the amygdalin single-use group. Therefore, compared with the spironolactone single-use group and the amygdalin single-use group, the pharmaceutical composition group of the present invention has better clinical therapeutic effects of promoting blood circulation, removing blood stasis, reducing swelling and relieving pain.

[0064] Example 4 Effect of the pharmaceutical composition of the present invention on acute soft tissue injury in rats

[0065] 1. Experimental Methods

[0066] 32 clean SD rats, female, weighing 180-200 g, with free access to water and food, and the room temperature was maintained at 20-25°C.

[0067] 24 hours before modeling, the hair on the rats' hind legs and thighs was removed with 8% sodium sulfide depilatory agent, and the rat model of acute soft tissue injury was established by local percussion.

[0068] After modeling, the rats were randomly divided into 4 groups: spironolactone group, amygdalin group, combination group, and model group, with 8 rats in each group. Each group of rats was gavaged, with 0.6 mg / kg spironolactone per rat in the spironolactone group, 6.0 mg / kg amygdalin per rat in the amygdalin group, 0.3 mg / kg spironolactone + 3.0 mg / kg amygdalin per rat in the combination group, and 0.2 mL normal saline per rat in the model group. The drugs were administered for 7 consecutive days, once a day. Blood was collected from each group of animals on the morning of the 7th day to measure blood rheology indicators (whole blood viscosity and plasma viscosity).

[0069] One hour after the last administration, the rats in each group were anesthetized and fixed on the operating board to collect blood from the abdominal aorta. After the blood collection was completed, the blood samples were centrifuged and the upper serum was collected. The serum samples were collected into the marked EP tubes with a pipette and stored in a -80℃ refrigerator for backup. The ELISA method was used to detect the content of PGE2 and IL-6 in the serum. The specific detection steps were performed according to the instructions on the kit.

[0070] 2. Statistical methods

[0071] Graphpad Prism 9.0 software was used for data processing, and the measured data were expressed as mean ± standard deviation. One-way analysis of variance was used for comparison among multiple groups, and SNK-q was used for pairwise comparison among groups. The difference was considered statistically significant when P < 0.05.

[0072] 3. Experimental results

[0073] 3.1 Effects on whole blood viscosity and plasma viscosity in rats

[0074] Compared with the model group, the combination group significantly reduced the whole blood viscosity and plasma viscosity in the rat serum, and its effect of reducing the expression of PGE2 in the rat serum, whole blood viscosity and plasma viscosity was significantly better than that of the spironolactone alone group and the amygdalin alone group. Figure 4 and Figure 5 .

[0075] 3.2 Effects on rat serum PGE2

[0076] Compared with the model group, the combination group significantly reduced the expression level of PGE2 in rat serum, and its effect of reducing the expression level of PGE2 in rat serum was significantly better than that of the spironolactone alone group and the amygdalin alone group.

[0077] The pharmaceutical composition of the present invention can significantly reduce the expression of PGE2 in rat serum. The two active ingredients, spironolactone and amygdalin, show significant synergistic effects in reducing the expression of PGE2 in rat serum. Figure 6 .

[0078] 3.3 Effect on rat serum IL-6

[0079] Compared with the model group, the combination group significantly reduced the expression level of IL-6 in rat serum, and its effect of reducing the expression level of IL-6 in rat serum was significantly better than that of the spironolactone alone group and the amygdalin alone group.

[0080] The pharmaceutical composition of the present invention can significantly reduce the expression of IL-6 in rat serum. The two active ingredients, spironolactone and amygdalin, show significant synergistic effects in reducing the expression of IL-6 in rat serum. Figure 7 .

[0081] The pharmaceutical composition of the present invention effectively improves the high viscosity, high polymerization and high concentration state of blood during blood stasis, relieves the blood stasis state, and thus achieves the function of promoting blood circulation and removing blood stasis.

[0082] The pharmaceutical composition of the present invention reduces the expression of PGE2 in rat serum, alleviates inflammatory symptoms such as swelling, fever, and pain in local tissues, and thus contributes to the recovery of acute soft tissue injuries; the pharmaceutical composition of the present invention reduces the expression of IL-6 in rat serum, reduces the inflammatory response, and thus accelerates the repair process of muscle tissue.

[0083] The above embodiments are merely examples for 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, and the obvious changes or modifications derived therefrom are still within the scope of protection of the present invention.

Claims

1. A pharmaceutical composition for promoting blood circulation, reducing swelling and relieving pain, characterized in that: The pharmaceutical composition consists of spironolactone and amygdalin, and the weight ratio of spironolactone to amygdalin is 1:8-12.

2. The pharmaceutical composition for promoting blood circulation, reducing swelling and relieving pain according to claim 1, characterized in that: The weight ratio of spironolactone to amygdalin in the pharmaceutical composition is 1:

10.

3. A drug preparation for promoting blood circulation, reducing swelling and relieving pain, characterized in that: The pharmaceutical preparation is prepared by mixing the pharmaceutical composition according to any one of claims 1 or 2 with pharmaceutically acceptable excipients.

4. The pharmaceutical preparation for promoting blood circulation, reducing swelling and relieving pain as claimed in claim 3, characterized in that: The pharmaceutical preparation is an oral preparation or an injection.

5. The pharmaceutical preparation for promoting blood circulation, reducing swelling and relieving pain according to claim 4, characterized in that: The oral preparation is powder, tablet, pill, capsule, granule or oral liquid.

6. Use of the blood-activating, swell-removing, and analgesic pharmaceutical composition according to any one of claims 1 or 2 or the blood-activating, swell-removing, and analgesic pharmaceutical preparation according to claim 3 in the preparation of blood-activating, stasis-removing, and swell-removing analgesic drugs.

7. Use of the blood-activating, swelling-reducing and analgesic pharmaceutical composition according to any one of claims 1 or 2 or the blood-activating, swelling-reducing and analgesic pharmaceutical preparation according to claim 3 in the preparation of drugs for treating traumatic injuries, tendon injuries and fractures, and soft tissue injuries.

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