“Pharmacological Activity & Manufacturing Method of 4-Medicinal-Plant-Derived Coronary Blood Flow Enhancing Agent” can be grouped into the US combined patent on pharmacology, isolation and purification of medicinal plants. This agent can increase coronary blood flow, decrease heart rate, low blood pressure and left ventricular pressure, reduce oxygen consumption index and myocardial oxygen consumption in the isolated guinea pig heart perfusion model.
According to the Examination Guideline of US patent & Trademark Office, data from in vitro or animal experiment, in general, are sufficient to support the therapeutic practicality. If they are reasonably associated with specific therapy or pharmacological practicality, data from in vitro, animal or combined experiment are absolutely sufficient to determine the practicality of a certain compound, synthesized chemical or processing method. The evidences needn't be obtained from experiments with well-accepted related animal model. The experimental data, if they have reasonable relationship with the practicality described in the patent application, should be evaluated substantially. So, the data provided by the applier can be experimental report based on a specific animal model and combined with the appropriate explanation of the practicality supported by experiments. If technicians believe that the conclusion from animal experiment has reasonable prediction in the human practicality, considerable thinking should be given to the evidences from experiments which support the practicality. The Pharmacological Activity & Manufacturing Method of 4-Medicinal-Plant-Derived Coronary Blood Flow Enhancing Agent is consistent with the rules of patentability mentioned above. Physiologically, coronary artery supplies the myocardium with blood and oxygen. Although the heart only account fro 0.5% of the body weight, but the coronary blood flow volume is as high as 5% of the cardiac output. Not only the myocardial blood flow but also the blood oxygen uptake rate is high In the normal condition, the arterial oxygen content is 20%, but the venous oxygen content in the coronary sinus is only 7%, indicating that 70% oxygen was uptaken by the myocardium which is three times as high as that of the other organ (22%). It is because the aerobic metabolism predominates in the myocardium. Coronary blood flow is determined by myocardial oxygen consumption, so the determinant factors of myocardial oxygen consumption, so the determinant factors of myocardial oxygen consumption also affects coronary blood flow. Normally speaking, if the heart beats stronger and faster, it needs more oxygen. So, the coronary blood flow will increase correspondingly to meet the increased oxygen demand. The major etiologic factor of myocardial ischemia or coronary heart disease is atherosclerosis which is related to low density lipoprotein, hyperhomocysteinemia, estrogen, and myocardial infarction. The classic drug for myocardial ischmia is nitroglycerine, which takes effect through providing exogenous nitrogen oxide (NO). The 4-Medicinal-Plant-Derived Coronary Blood Flow Enhancing Agent is more suitable for the prevention and preventive therapy of myocardial ischemia.
Both the left and right coronary artery originate from the aoritic root and supply the myocardium with nutrient and oxygen. In the isolated heart perfusion experiment, the myocardium was perfused with oxygenated, thermostatic and pressure-constant KH solution via the coronary artery connected with aortic cannula; and the coronary venous effluence was collected for the coronary blood flow determination. Pharmacological experiment indicated that the 4-Medicinal-Plant-Derived Coronary Blood Flow Enhancing Agent increased the coronary blood flow in the isolated guinea pig heart. Isoprenaline obviously increased heart rate and myocardial contraction although it enhanced the coronary blood flow.
In the pharmacological experiment, the hemodynamic and coronary blood flow of the anaesthetized canine was studied by Eight Channel Physiological Recorder. The results indicated that this agent slowed down the heart rate, lowered the blood pressure, decreased the left ventricular pressure and reduced the oxygen consumption index as well as the myocardial oxygen consumption. This agent also protected the ischemic myocardium and reduced the myocardial ischemia degree by lowering the peripheral vascular resistance, regulating the oxygen supply and demand balance, decreasing the myocardial tension and improving the blood supply of the endocardium. In addition, this agent improved the cardiac function and hemodynamics by dilating coronary artery, increasing coronary blood flow, decreasing myocardial oxygen consumption, regulating myocardial metabolism and lowering the peripheral vascular resistance. This agent is useful in the prevention and preventive therapy of myocardial ischemia, coronary heart disease and angina pectoris. (Table 3).
Rhizoma Chunanxiong
Radix Salviae Miltiorrhizae
Fios Carthami
Rhizoma Corydalis
Part III Pharmacological Activity of 4-Medicinal-Plant-Derived Coronary Blood Flow Enhancing Agent
1. Effect on coronary blood flow of the isolated guinea pig heart.
Results: This agent increased the coronary blood flow of the isolated guinea pig heart. Although it increased the coronary blood flow, isoprenaline increased the heart rate and myocardial contraction. (Table 1)
2. Effect on the Hemodynamics and Coronary Blood Flow of Anaesthetized Canine. Dogs, weighing 15˜20 kg, were anaesthetized by intravenous injection of sodium pentobarbital (30 mg/kg), and fixed on the operating table in the right lateral decubitus position. The preparations included: Shaving the neck, chest an inner hind leg; isolating femoral vein and inserting venous cannula; mounting the microsyringe (first slowly transfusing normal saline); isolating trachea for artificial ventilation; seperating carotid and threading two suturing thread under the carotid for cannulation. The thoracectomy was performed along the 4˜5 intercostal space and the heart was exposed. Positive artificial ventilation was started (15˜20 cycles/min). Pericardium was opened longitudinally to make the “pericardial cradle”. On the top of the isolated aortic root and left coronary anterior descending branch (or left circumflex branch), the electromagnetic flowmeter probe was placed to measure cardiac output and coronary blood flow (10˜15 mm calibred probe for aorta; 1.5˜3 mm calibred probe for coronary artery.). The heparinized-perftisate-filled catheters were inserted into common carotid and left ventricle to measure the blood pressure and intraventricular pressure respectively, 5 mg/kg heparin was injected via femoral vein though T-Value. The ECG electrode was inserted under the skin of the extremities to record the lead II electrocardiograph and count the heart rate. The electrical signal of the left intraventricular pressure was input into the electrical differential analyzer to measure the left intraventricular pressure change rate. All date were input into the Eight Channel Physiological Recorder Simultaneously.
The normal graph was recorded first, and then recorded again intravenous injection of test sample via femoral vein. (See Table 2)
All the data mentioned above were primary parameters, from which the second parameters of hemodynamics were deduced.
Results: This agent slowed down the normal animals heart rate, lowered blood pressure as well as left intraventricular pressure, and decreased the oxygen consumption index and myocardial oxygen consumption.
1. “The Pharmacological Activity & Manufacturing Method of the 4-Medicinal-Plant-Derived Coronary Blood Flow Enhancing Agent” can be grouped in the US combined patent on pharmacology, isolation and purification of medicinal plant. The formula consists of Rhizoma Chuanxiong 20%, Radix Salviae Miltiorrhizae 33%, Fios Carthami 20%, Rhizoma Corydalis 27%.
2. The processing method can be described as follows: extracting with Carbon Dioxide supercritical fluid and separating the extractant for rectification; Mixing 300 kg, 200 kg, 100 kg 65% ethanol with 100 kg raw material and reflux extracting at 85° C. for 3, 2, 1 hours respectively; Concentrating the extractant and adding 100 kg water to 100 kg concentrate; staying rest for sendimentation at room temperature for 6 hours; collecting supernatant and adsorbing with WLD resin column; eluting with ethanol; collecting elution and extractant of carbon Dioxide Supercritical fluid extraction; Spraying-dry at 85° C.; high-performance liquid chromatography and fingerprint analysis to ensure the quality.
3. Pharmacological experiments indicated that this agent increased the coronary blood flow of the isolated guinea pig heart. The further experiment was conducted to study the hemodynamics and coronary blood flow in the anaesthetized canine model, and the results indicated this agent slowed down heart rate, lowered blood pressure, decreased left intraventricular pressure and reduced the oxygen consumption index oxygen consumption.