Use of bradykinin in the preparation of a medicament for reducing food intake, pharmaceutical composition

Abstract

The present invention provides an application of bradykinin in the preparation of a drug for reducing food intake, and a pharmaceutical composition, belonging to the technical field of biomedicine. The application of bradykinin or a pharmaceutically acceptable derivative thereof includes the application of bradykinin in the preparation of a drug for reducing food intake, and bradykinin has the amino acid sequence shown in SEQ ID No. 1. Bradykinin can inhibit food intake and reduce body weight through peripheral receptors rather than through the central nervous system; further, it can significantly reduce the food intake of mice and reduce the body weight of mice, which proves that bradykinin has the potential to treat obesity and can be used in the preparation of drugs for treating food intake inhibition or obesity. Moreover, bradykinin is an endogenous substance with high safety and relatively small side effects, and bradykinin acts on the peripheral system, avoiding the side effects of drugs acting on the central nervous system.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and particularly to the use of bradykinin as a drug for reducing food intake and a pharmaceutical composition. Background Art

[0002] There is a close pathophysiological link between obesity and various metabolic diseases, among which the most prominent are type 2 diabetes, hypertension, coronary atherosclerotic heart disease and other cardiovascular diseases. In addition, obesity may also cause various complications such as sleep apnea syndrome, osteoarthritis, non-alcoholic fatty liver disease, etc., seriously affecting the quality of life and life expectancy of patients.

[0003] At present, the clinical treatment of obesity mainly adopts a multi-modal comprehensive intervention strategy, including lifestyle interventions such as diet control and exercise therapy, drug treatment, and surgery. Among them, lifestyle intervention, as a basic treatment method, is safe but has limited effects and is often difficult to adhere to in the long term; surgical interventions such as gastric bypass surgery, although effective, have high risks and high costs, and are only applicable to severely obese patients. In terms of drug treatment, related drugs such as orlistat and liraglutide have certain curative effects, but generally have side effects such as gastrointestinal adverse reactions and cardiovascular risks, and the long-term treatment effects are limited. Therefore, it is necessary to develop new anti-obesity drugs with better safety, more significant weight loss effects and multiple benefits of improving metabolic indicators. Summary of the Invention

[0004] In view of this, in order to at least partially solve the above-mentioned technical problems, the present invention provides the use of bradykinin as a drug for reducing food intake and a pharmaceutical composition.

[0005] According to an embodiment of the present invention, there is provided the use of bradykinin or a pharmaceutically acceptable derivative thereof, the use including the use of bradykinin for preparing a drug for reducing food intake, and bradykinin has the amino acid sequence shown in SEQ ID No.1.

[0006] According to an embodiment of the present invention, bradykinin inhibits food intake and reduces food intake through peripheral receptors.

[0007] According to an embodiment of the present invention, the use further includes the use of bradykinin as a drug for treating obesity and / or reducing body weight.

[0008] According to an embodiment of the present invention, the use includes salifying bradykinin and / or chemically modifying the amino acid sequence shown in SEQ ID No.1.

[0009] According to an embodiment of the present invention, the salts formed by bradykinin include at least one of metal salts of bradykinin, salts formed by bradykinin and inorganic acids, and salts formed by bradykinin and organic acids; the chemical modifications in the chemical modification products of the amino acid sequence shown in SEQ ID No.1 include at least one of acetylation, amination, methylation, phosphorylation, glycosylation, lipidation, ubiquitination, biotin labeling, and fluorescent protein labeling.

[0010] According to an embodiment of the present invention, the metal salts of bradykinin include at least one of lithium salt, sodium salt, potassium salt, calcium salt, magnesium salt, manganese salt, copper salt, zinc salt, and aluminum salt; the inorganic acids include hydrochloric acid, sulfuric acid, boric acid, or carbonic acid; the organic acids include at least one of acetic acid, citric acid, lactic acid, malonic acid, maleic acid, tartaric acid, fumaric acid, benzoic acid, aspartic acid, glutamic acid, succinic acid, oleic acid, trifluoroacetic acid, and oxalic acid.

[0011] According to an embodiment of one aspect of the present invention, a pharmaceutical composition is provided, and the pharmaceutical composition is used for reducing food intake and / or treating obesity; the pharmaceutical composition includes bradykinin having the amino acid sequence shown in SEQ ID No.1 or a pharmaceutically acceptable derivative thereof as an active ingredient.

[0012] According to an embodiment of the present invention, the derivatives include salts of bradykinin and / or chemical modification products of bradykinin.

[0013] According to an embodiment of the present invention, the dosage form of the pharmaceutical composition is any one of aerosol, spray, powder, pill, tablet, film, ointment, suppository, paste, solution, injection, mixture, lotion, or liniment.

[0014] According to an embodiment of the present invention, the dosage of the pharmaceutical composition is 0.1 - 100 mg / kg.

[0015] According to an embodiment of the present invention, it is proposed that bradykinin can inhibit food intake and reduce body weight through peripheral receptors rather than through the central nervous system; further, it can significantly reduce the food intake of mice and reduce the body weight of mice, which can prove that bradykinin has the potential to treat obesity, can be used in the preparation of drugs for treating food intake inhibition or obesity, and bradykinin is an endogenous substance with high safety, relatively small side effects, and bradykinin acts on the peripheral system, avoiding the side effects of central nervous system drugs. Description of the Drawings

[0016] Figure 1 It is a comparative diagram of the results of the influence of intraperitoneal injection of bradykinin and phosphate buffered saline (PBS) on the food intake of mice in Example 1 of the present invention;

[0017] Figure 2Comparison chart of the effects of intraperitoneal injection of bradykinin and PBS on mouse body weight in Example 1 of the present invention;

[0018] Figure 3 Comparison chart of the effects of intracerebroventricular injection of bradykinin and PBS on mouse food intake in Example 2 of the present invention;

[0019] Figure 4 Comparison chart of the effects of intracerebroventricular injection of bradykinin and PBS on mouse body weight in Example 2 of the present invention. Detailed implementation manners

[0020] Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. However, it should be understood that these descriptions are merely exemplary and are not intended to limit the scope of the present invention. In the following detailed description, for the sake of explanation, many specific details are set forth in order to provide a comprehensive understanding of the embodiments of the present invention. However, obviously, one or more embodiments can also be implemented without these specific details. In addition, in the following description, descriptions of well-known structures and technologies are omitted to avoid unnecessarily confusing the concepts of the present invention.

[0021] The terms used herein are merely for describing specific embodiments and are not intended to limit the present invention. The term "comprising" used herein indicates the presence of features, steps, operations, but does not exclude the presence or addition of one or more other features.

[0022] In the present invention, the term "pharmaceutically acceptable" refers to a compound, substance, composition, and / or dosage form that is applicable to contact with tissues of humans and animals within the scope of reasonable medical judgment without excessive toxicity, irritation, allergic reaction, or other problems or complications, and is commensurate with a reasonable benefit / risk ratio.

[0023] The term "treatment" means that after a subject suffers from a disease, the subject is contacted (such as administered) with a drug, composition, etc. based on the present invention, so that the symptoms of the disease are alleviated compared with the situation without contact, and it does not necessarily mean that the symptoms of the disease are completely inhibited. Suffering from a disease means that the body shows symptoms of the disease.

[0024] In the process of realizing the concept of the present invention, it is found that bradykinin is a bioactive peptide composed of 9 amino acids, and mainly plays a key role in various physiological and pathological processes by activating two types of G protein-coupled receptors (GPCRs), namely B1 and B2. Previous studies have shown that it has important functions in inflammatory responses, vasodilation, blood pressure regulation, pain perception, etc., but its potential in the treatment of obesity has not been explored.

[0025] Specifically, according to an embodiment of an aspect of the present invention, there is provided an application of bradykinin or a pharmaceutically acceptable derivative thereof, the application including the use of bradykinin for preparing a drug for reducing food intake, and bradykinin has the amino acid sequence shown in SEQ ID No. 1.

[0026] According to an embodiment of the present invention, it is proposed that bradykinin can inhibit food intake and reduce body weight through peripheral receptors rather than through the central nervous system; further, it can significantly reduce the food intake of mice and reduce the body weight of mice, which can prove that bradykinin has the potential to treat obesity and can be used in the preparation of drugs for treating food intake inhibition or obesity. Moreover, bradykinin is an endogenous substance with high safety and relatively small side effects, and bradykinin acts on the peripheral system, avoiding the side effects of central nervous system drugs.

[0027] According to an embodiment of the present invention, the amino acid sequence shown in SEQ ID No. 1 is: RPPGFSPFR, or Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg, which is a linear nonapeptide.

[0028] According to an embodiment of the present invention, bradykinin inhibits food intake and reduces food intake through peripheral receptors.

[0029] According to an embodiment of the present invention, bradykinin binds to B1 and B2 receptors in peripheral tissues such as the gastrointestinal tract or blood vessels, regulates appetite-related signaling pathways, and affects the secretion of gastrointestinal hormones such as cholecystokinin and glucagon-like peptide-1 (GLP-1), thereby reducing hunger or enhancing satiety, and finally achieving the effect of reducing food intake. This mechanism avoids the possible side effects of directly acting on the central nervous system and has higher safety and controllability.

[0030] According to an embodiment of the present invention, the application further includes the use of bradykinin as a preparation for treating obesity and / or reducing body weight drugs.

[0031] According to an embodiment of the present invention, by reducing food intake and suppressing appetite, bradykinin can effectively reduce energy intake, thereby helping to control body weight and treat obesity.

[0032] According to an embodiment of the present invention, the application includes salifying bradykinin and / or chemically modifying the amino acid sequence shown in SEQ ID No. 1.

[0033] According to an embodiment of the present invention, by salifying or chemically modifying the amino acid sequence of bradykinin, its stability, bioavailability or drug efficacy can be improved. Salification and chemical modification can improve the pharmacokinetic properties of bradykinin, such as prolonging the half-life, enhancing targeting or reducing degradation, thereby enhancing its effect and convenience in practical applications.

[0034] According to an embodiment of the present invention, the salts formed from bradykinin include at least one of metal salts of bradykinin, salts formed from bradykinin and inorganic acids, and salts formed from bradykinin and organic acids; the chemical modifications in the chemical modifications of the amino acid sequence shown in SEQ ID No.1 include at least one of acetylation, amination, methylation, phosphorylation, glycosylation, lipidation, ubiquitination, biotinylation, and fluorescent protein labeling.

[0035] According to an embodiment of the present invention, the formed salts refer to a salt recognized for use in animals, and more precisely in humans, including metal salts of the amino acid sequence shown in SEQ ID No.1, and the metals include but are not limited to: lithium, sodium, potassium, calcium, magnesium, manganese, copper, zinc, or aluminum, etc.; including salts formed from the amino acid sequence shown in SEQ ID No.1 and organic bases, and the organic bases include but are not limited to: ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, arginine, lysine, histidine, or piperazine, etc.; including salts formed from the amino acid sequence shown in SEQ ID No.1 and inorganic acids or organic acids, and the organic acids include but are not limited to: acetic acid, citric acid, lactic acid, malonic acid, maleic acid, tartaric acid, fumaric acid, benzoic acid, aspartic acid, glutamic acid, succinic acid, oleic acid, trifluoroacetic acid, oxalic acid, pamoate, or gluconic acid, etc.; the inorganic acids include but are not limited to: hydrochloric acid, sulfuric acid, boric acid, or carbonic acid.

[0036] According to an embodiment of another aspect of the present invention, a pharmaceutical composition is provided, and the pharmaceutical composition is used to reduce food intake and / or treat obesity; the pharmaceutical composition includes bradykinin having the amino acid sequence shown in SEQ ID No.1 or a pharmaceutically acceptable derivative thereof as an active ingredient.

[0037] According to an embodiment of the present invention, the pharmaceutical composition further includes bradykinin and its pharmaceutically acceptable derivatives.

[0038] According to an embodiment of the present invention, when the drug active ingredient is a pharmaceutically acceptable salt, such as a pharmaceutically acceptable acid addition salt or base addition salt, it can react with a pH regulator in a solution to thereby in-situ prepare the corresponding free active ingredient, such as a free basic active ingredient or a free acidic active ingredient.

[0039] According to an embodiment of the present invention, the dosage form of the pharmaceutical composition is any one of aerosol, spray, powder, pill, tablet, film, ointment, suppository, paste, solution, injection, mixture, lotion, or liniment.

[0040] According to the embodiments of the present invention, the dosage forms of various drugs can meet different treatment needs, improve patient compliance, optimize the efficacy and safety of drugs, meet different clinical needs and patient preferences, and adapt to different administration routes.

[0041] Specifically, it may further include adjuvant components, including at least one of preservatives, solubilizers, stabilizers, diluents, and lubricants. The adjuvant components can improve the physicochemical properties of the drugs, enhance the efficacy and safety of the drugs, increase the acceptability of the preparations, meet different clinical needs and patient preferences, and at the same time contribute to the stable release and effective delivery of the drugs, and also improve the safety of the preparations. By reasonably selecting and using these excipients, the dosage forms of the drugs can be optimized and the therapeutic effect can be improved.

[0042] According to the embodiments of the present invention, the dosage of the pharmaceutical composition is 0.1 - 100 mg / kg.

[0043] According to the embodiments of the present invention, the dosage can be 0.1 mg / kg, 10 mg / kg, 20 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, but is not limited to the listed values, and other unlisted values within this numerical range are equally applicable.

[0044] According to the embodiments of the present invention, the administration method is preferably by intraperitoneal injection, but is not limited to only intraperitoneal injection, and other methods such as oral administration can be adjusted with appropriate dosage forms.

[0045] The following will further explain and illustrate the solution of the present invention with specific examples. In the following examples, if not otherwise specified, all are conventional commercially available reagents.

[0046] In the following result graphs of the examples, * represents P value < 0.05, ** represents P value < 0.01, *** represents P value < 0.001, and only when the P value < 0.05 is the data considered to have a statistically significant difference. The data values in the statistical result graphs are represented by the mean (n is the total number of experimental groups) ± standard error.

[0047] Example 1 Effect of Intraperitoneal Injection of Bradykinin on Diet and Body Weight of Mice

[0048] Experimental animals: Eight-week-old male C57BL / 6J mice with normal development were selected as experimental subjects and divided into an experimental group and a control group. Before the formal experiment, the mice were individually housed for 3 days to adapt.

[0049] Drug preparation: Bradykinin was dissolved in PBS buffer to prepare a solution with an appropriate concentration, and it was prepared freshly for immediate use.

[0050] Intraperitoneal injection experiment: After fasting the two groups of mice overnight for 12 hours, fix the mice. Use a 1 ml syringe to draw an appropriate amount of bradykinin solution and slowly inject it into the peritoneal cavity at the lower 1 / 3 of the mouse abdomen, avoiding the internal organs. The dose of bradykinin for intraperitoneal injection is 10 mg / kg, and the control group is injected with an equal amount of PBS buffer. The operation steps are the same as those of the experimental group.

[0051] Monitor the food intake of the mice within 5 hours, that is, monitor the food intake at 0, 0.5, 1, 1.5, 2, and 5 h. The results are as Figure 1 shown. Inject bradykinin or PBS at the same time point every day for one week, and record the body weight of the mice at the same time point before injecting the drug every day. The results are as Figure 2 shown.

[0052] Figure 1 This is a comparative graph of the effects of intraperitoneal injection of bradykinin and PBS on the food intake of mice in Example 1 of the present invention; Figure 2 This is a comparative graph of the effects of intraperitoneal injection of bradykinin and PBS on the body weight of mice in Example 1 of the present invention.

[0053] According to Figure 1 and Figure 2 it can be seen that after intraperitoneal injection of bradykinin, the food intake of the mice decreased significantly, and after continuous injection for one week, the body weight of the mice showed a downward trend.

[0054] Example 2 Effects of intracerebroventricular injection of bradykinin on the diet and body weight of mice

[0055] Experimental animals: Select 8-week-old male C57BL / 6J mice with normal development as experimental subjects and divide them into an experimental group and a control group. Before the formal experiment, the mice are individually housed and adapted for 3 days.

[0056] Drug preparation: Dissolve bradykinin in PBS buffer to prepare a solution with an appropriate concentration, and prepare it for immediate use.

[0057] Intracerebroventricular injection experiment: For the mice in the experimental group and the control group, cannulas are implanted into the lateral ventricles through a stereotaxic apparatus, and the experimental operation is carried out after the mice recover for one week. After fasting the two groups of mice overnight for 12 hours, bradykinin is injected into the lateral ventricle through the cannula with the aid of a microinjection pump. The dose of bradykinin for intracerebroventricular injection is 10 nM / 2 μl, and the control group is injected with an equal amount of PBS buffer. The operation steps are the same as those of the experimental group.

[0058] Monitor the food intake of the mice within 5 hours after intracerebroventricular injection of bradykinin, that is, monitor the food intake at 0, 0.5, 1, 1.5, 2, and 5 h. The results are as Figure 3 shown. Inject bradykinin or PBS at the same time point every day for one week, and record the body weight of the mice at the same time point before injecting the drug every day. The results are asFigure 4 as shown

[0059] Figure 3 It is a comparison chart of the effects of bradykinin and PBS injected into the lateral ventricle on the food intake of mice in Example 2 of the present invention; Figure 4 It is a comparison chart of the effects of bradykinin and PBS injected into the lateral ventricle on the body weight of mice in Example 2 of the present invention.

[0060] According to Figure 3 and Figure 4 it can be seen that after injecting bradykinin into the lateral ventricle, the food intake of the mice did not change significantly, and after continuous injection for one week, there was also no obvious effect on the body weight of the mice.

[0061] In summary, it can be seen that bradykinin inhibits food intake and reduces body weight through peripheral receptors, rather than acting through the central nervous system.

[0062] The specific embodiments described above further elaborate on the purpose, technical solutions, and beneficial effects of the present invention. It should be understood that the above are only specific embodiments of the present invention and are not used to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims

1. Use of bradykinin or a pharmaceutically acceptable derivative thereof, characterized in that, it includes use in the preparation of a drug for reducing food intake, and the bradykinin has the amino acid sequence shown in SEQ ID No.

1.

2. Use of bradykinin or a pharmaceutically acceptable derivative thereof, characterized in that, it includes use in the preparation of a drug for treating obesity, and the bradykinin has the amino acid sequence shown in SEQ ID No.

1.

3. The use according to claim 1 or 2, characterized in that, the use includes salifying the bradykinin.

4. The use according to claim 1 or 2, characterized in that, the salt formed from the bradykinin includes at least one of a metal salt of bradykinin, a salt formed from bradykinin and an inorganic acid, and a salt formed from bradykinin and an organic acid.

5. The use according to claim 4, characterized in that, the metal salt of the bradykinin includes at least one of lithium salt, sodium salt, potassium salt, calcium salt, magnesium salt, manganese salt, copper salt, zinc salt, and aluminum salt.

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