Heme composition, pharmaceutical composition and application thereof
By adding superoxide dismutase and catalase to heme, a heme composition is formed, which solves the problems of instability and low utilization of existing iron supplements and achieves high stability and low toxicity of heme. Side effects, suitable for long-term use.
Patent Information
- Application Number
- CN202111129448.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2021-09-26
- Publication Date
- 2025-01-21
- Estimated Expiration
- 2041-09-26
AI Technical Summary
The existing iron supplements have low utilization rates in the body, great toxic side effects and unstable, and are difficult to take for a long time, and cannot effectively prevent the heme iron in heme from being oxidized and decomposed.
A heme composition is provided, including heme, superoxide dismutase and catalase, which can effectively prevent the oxidative decomposition of heme iron and improve the stability of heme through mixing in a reasonable proportion.
By adding superoxide dismutase and catalase, the stability of heme is significantly improved, its utilization in the body is enhanced, and toxic and side effects are reduced, making it suitable for long-term use.
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Abstract
Description
Technical Field
[0001] The present application relates to the field of hemoglobin technology, and in particular to a hemoglobin composition, a pharmaceutical composition and applications thereof. Background Art
[0002] Iron is an essential nutrient for the human body. It is mainly found in hemoglobin and myoglobin. It is responsible for transporting oxygen, catalyzing the oxidation of organic matter, decomposing peroxides and transferring electrons in the blood. At present, the prevention and treatment of iron deficiency anemia is mainly achieved through iron supplements. Common iron supplements include ferrous sulfate, ferrous chloride, ferrous gluconate, ferrous lactate, ferrous succinate and ferrous fumarate. Although these iron supplements have high iron content and good iron supplementation effects, they are difficult to take for a long time because of their low utilization rate in the body, large toxic side effects, and a special metallic rust taste.
[0003] Heme iron in heme is an organic iron that can be directly absorbed by intestinal mucosal cells without causing digestive tract irritation symptoms and has a high bioavailability. However, heme iron is unstable and easily oxidized and decomposed, thus affecting its stability. Summary of the invention
[0004] The purpose of the present application is to provide a heme composition that can effectively prevent the heme in hemoglobin from being oxidized and decomposed, thereby improving its stability.
[0005] In a first aspect, the present application provides a heme composition, comprising: heme, superoxide dismutase and catalase.
[0006] In some embodiments, the ratio of the mass of the heme, the enzyme activity of the superoxide dismutase, and the enzyme activity of the catalase is 5 g: 3000-6000 IU: 1000-3000 IU.
[0007] The second aspect of the present application provides the use of the heme composition provided in the first aspect of the present application in the preparation of a medicament for preventing or treating iron deficiency-related diseases.
[0008] In some embodiments, the iron deficiency-related disease comprises iron deficiency anemia.
[0009] The third aspect of the present application provides a pharmaceutical composition, comprising the heme composition provided in the first aspect of the present application and a pharmaceutically acceptable carrier, diluent or excipient.
[0010] In some embodiments, the dosage form is an oral dosage form.
[0011] In some embodiments, the oral dosage form is selected from one of oral liquid, syrup, granules, capsules, tablets, powders, pills and pellets.
[0012] The fourth aspect of the present application provides the use of the pharmaceutical composition provided in the third aspect of the present application in the preparation of a medicament for preventing or treating iron deficiency-related diseases.
[0013] In some embodiments, the iron deficiency-related disease comprises iron deficiency anemia.
[0014] The heme composition provided in the present application can effectively prevent heme from being oxidized and decomposed by adding superoxide dismutase and catalase to heme, thereby improving the stability of heme.
[0015] Of course, implementing any product or method of the present application does not necessarily require achieving all of the advantages described above at the same time. DETAILED DESCRIPTION
[0016] The technical solutions in this application are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of this application, not all of them. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field based on this application belong to the scope of protection of this application.
[0017] In a first aspect, the present application provides a heme composition, comprising: heme, superoxide dismutase and catalase.
[0018] Superoxide dismutase is an important antioxidant enzyme in the body, which can remove excess oxygen free radicals produced during the body's metabolism and delay the aging of the body. Catalase is one of the key enzymes in the biological defense system established during the biological evolution process. Its biological function is to catalyze the decomposition of hydrogen peroxide in cells and prevent delipidation.
[0019] The inventors found in the study that mixing heme, superoxide dismutase and catalase can effectively prevent the low-valent iron in heme from being oxidized to high-valent iron, thereby improving the stability of heme; only mixing heme and superoxide dismutase, or only mixing heme and catalase, is difficult to effectively prevent the low-valent iron in heme from being oxidized to high-valent iron. In addition, the present application mixes heme with superoxide dismutase and catalase, so that the obtained heme composition has both the function of iron supplementation and the functions of anti-oxidation, beauty and anti-aging.
[0020] In some embodiments of the present application, the ratio of the mass of heme, the enzyme activity of superoxide dismutase, and the enzyme activity of catalase is 5 g: 3000-6000 IU: 1000-3000 IU.
[0021] The inventors found in their research that mixing heme, superoxide dismutase and catalase in the above ratio can effectively prevent the low-valent iron in heme from being oxidized into high-valent iron, thereby improving the stability of heme.
[0022] The second aspect of the present application provides the use of the heme composition provided in the first aspect of the present application in the preparation of a medicament for preventing or treating iron deficiency-related diseases.
[0023] In some embodiments of the present application, the iron deficiency-related disease includes iron deficiency anemia.
[0024] The iron deficiency anemia described in the present application can be selected from one or more of iron deficiency anemia in pregnant women, latent iron deficiency anemia in children and adolescents, iron deficiency anemia caused by gastrointestinal disorders, iron deficiency anemia caused by blood loss, iron deficiency anemia caused by menstruation, iron deficiency anemia caused by injury, iron deficiency anemia caused by sprue, iron deficiency anemia in cancer patients, iron deficiency anemia caused by chemotherapy, iron deficiency anemia caused by inflammation, iron deficiency anemia caused by chronic heart failure, iron deficiency anemia caused by rheumatoid arthritis, iron deficiency anemia caused by systemic lupus erythematosus and iron deficiency anemia caused by inflammatory bowel disease.
[0025] The third aspect of the present application provides a pharmaceutical composition, comprising the heme composition provided in the first aspect of the present application and a pharmaceutically acceptable carrier, diluent or excipient.
[0026] The pharmaceutically acceptable carrier, diluent or excipient in the pharmaceutical composition of the present application can be any conventional carrier, diluent or excipient in the field of pharmaceutical preparations, and the selection of a specific carrier, diluent or excipient depends on the mode of administration or the type of disease and state used to treat a specific patient. The preparation method of the pharmaceutical composition for a specific mode of administration is completely within the knowledge of the technical personnel in the field of medicine. For example, it can be used as a pharmaceutically acceptable carrier, diluent or excipient, including conventional starch, icing sugar, dextrin, lactose, compressible starch, starch slurry, sodium carboxymethyl cellulose, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, microcrystalline cellulose, hydroxypropyl methyl cellulose, vitamin C, lactose, magnesium stearate, micropowder silica gel, talc, hydrogenated vegetable oil, polyethylene glycols and magnesium lauryl sulfate, etc.
[0027] In some embodiments of the present application, the dosage form is an oral dosage form.
[0028] In some embodiments of the present application, the oral dosage form is selected from one of oral liquid, syrup, granules, capsules, tablets, powders, pills and pellets.
[0029] For different types of oral dosage forms, different carriers, diluents or excipients can be selected, which are conventional technical means in the art. For example, when preparing capsules, lactose, magnesium stearate, starch and powdered sugar can be used; when preparing tablets, sodium carboxymethyl cellulose and dextrin can be used.
[0030] The pharmaceutical compositions in various dosage forms mentioned above can be prepared according to conventional methods in the pharmaceutical field.
[0031] The fourth aspect of the present application provides the use of the pharmaceutical composition provided in the third aspect of the present application in the preparation of a medicament for preventing or treating iron deficiency-related diseases.
[0032] In some embodiments of the present application, the iron deficiency-related disease includes iron deficiency anemia.
[0033] The content of "iron deficiency anemia" described in the fourth aspect of this application is the same as the corresponding content of the above-mentioned second aspect, and will not be repeated here.
[0034] The terms used in this application, unless otherwise specified or defined, have their general meanings known to those skilled in the art.
[0035] As used herein, the term "treatment" has its general meaning, such as treating an individual who may suffer from an iron deficiency-related disease or is at risk of suffering from an iron deficiency-related disease with the pharmaceutical composition of the present application, in order to treat, cure, alleviate, or mitigate the disease. Similarly, as used herein, the term "prevention" has its general meaning, such as treating an individual who may suffer from an iron deficiency-related disease or is at risk of suffering from an iron deficiency-related disease with the pharmaceutical composition of the present application, in order to prevent, prevent, block, or isolate the disease.
[0036] As used in this application, the term "pharmaceutical composition" has its general meaning, and in this application may refer to a preparation further processed from the heme composition of this application, such as a pharmaceutical preparation. In addition, the "pharmaceutical composition" of this application may also be present or provided in the form of health products, functional foods, foods, food additives, etc. The pharmaceutical composition of this application may be prepared by conventional techniques in the pharmaceutical field, especially in the field of formulations, optionally mixed with one or more pharmaceutically acceptable carriers, diluents or excipients, and then made into the desired dosage form.
[0037] When the pharmaceutical compositions of the present application are administered to human or animal subjects, they can be administered by themselves, that is, without adding any of the above-mentioned pharmaceutically acceptable carriers, diluents or excipients, and the pharmaceutical compositions of the present application can be directly administered to the patient in their original form; or they can be administered to human or animal subjects after containing, for example, 1%-99% of the pharmaceutical composition or the pharmaceutical composition is combined with a pharmaceutically acceptable carrier, diluent or excipient.
[0038] The actual dosage level and administration schedule of the pharmaceutical composition or active ingredient in the pharmaceutical composition of the present application can be adaptively changed so that for a specific subject, composition and administration method, the amount of the active ingredient can effectively obtain the desired therapeutic response without being toxic to the subject.
[0039] The present application is described in detail below with reference to specific embodiments.
[0040] Example 1
[0041] 5 g of heme, 3000 IU of superoxide dismutase, and 1000 IU of catalase were mixed to obtain a heme composition.
[0042] Example 2
[0043] 5 g of heme, 6000 IU of superoxide dismutase and 3000 IU of catalase were mixed to obtain a heme composition.
[0044] Example 3
[0045] 5 g of heme, 4500 IU of superoxide dismutase and 2000 IU of catalase were mixed to obtain a heme composition.
[0046] Example 4
[0047] 5 g of heme, 1 g of sodium carboxymethylcellulose, 4500 IU of superoxide dismutase and 2000 IU of catalase were mixed to obtain a mixture.
[0048] 10 wt % of dextrin is added to the mixture to prepare a soft material, which is then tableted and dried to obtain tablets.
[0049] Example 5
[0050] 5 g of heme, 1 g of lactose, 0.5 g of magnesium stearate, 25 g of starch, 15 g of powdered sugar, 4500 IU of superoxide dismutase and 2000 IU of catalase were mixed to obtain a mixture.
[0051] The mixture is moistened with an ethanol aqueous solution with a volume fraction of 60% to form granules; the granules are dried and loaded into enteric-coated capsules to obtain capsules.
[0052] Example 6
[0053] The same procedures as in Example 5 were used except that the dosage of superoxide dismutase was 1000 IU and the dosage of catalase was 4000 IU.
[0054] Example 7
[0055] The same procedures as in Example 5 were used except that the dosage of superoxide dismutase was 7000 IU and the dosage of catalase was 500 IU.
[0056] Comparative Example 1
[0057] Except for not adding 4500 IU superoxide dismutase, the rest is the same as Example 5.
[0058] Comparative Example 2
[0059] Except that 2000 IU catalase was not added, the rest was the same as Example 5.
[0060] Comparative Example 3
[0061] The method is the same as Example 5 except that 2000 IU of catalase is not added and the amount of superoxide dismutase used is 35000 IU.
[0062] Comparative Example 4
[0063] The procedure was the same as in Example 5 except that 4500 IU superoxide dismutase and 2000 IU catalase were not added.
[0064] Comparative Example 5
[0065] Except that 4500 IU superoxide dismutase and 2000 IU catalase were not added, the rest was the same as Example 4.
[0066] Heme stability test
[0067] In order to determine the stability of hemoglobin in the products obtained in Examples 1-7 and Comparative Examples 1-5, the actual content of hemoglobin in the products obtained in Examples 1-7 and Comparative Examples 1-5 was tested by liquid chromatography.
[0068] 1. Test conditions
[0069] Chromatographic column: Agilent TC-C18 column (4.6 mm*250 mm, 5 μm).
[0070] Mobile phase: methanol: 0.6% acetic acid aqueous solution = 70:30 (volume ratio).
[0071] Flow rate: 1.0mL / min.
[0072] Column temperature: 35°C.
[0073] Injection volume: 20 μL.
[0074] Detection wavelength: 399nm.
[0075] 2. Prepare standard working solution
[0076] Accurately weigh 27.39 mg of hemin standard substance, dissolve it in 0.1 mol / L sodium hydroxide solution and make up to 50 mL, draw 1 mL, and make up to 25 mL with methanol.
[0077] Accurately pipette 0.1mL, 0.2mL, 0.3mL, 0.4mL, and 0.5mL of the above solution, dilute to 1mL with methanol, and prepare standard working solutions with concentrations of 2.1μg / mL, 4.2μg / mL, 6.3μg / mL, 8.4μg / mL, and 10.5μg / mL, respectively.
[0078] 3. Establish a standard curve
[0079] The standard working solutions were injected according to the liquid chromatography test conditions, and a standard curve was established with concentration as the horizontal axis and peak area as the vertical axis.
[0080] 4. Prepare the samples to be tested
[0081] After the products prepared in Examples 1-7 and Comparative Examples 1-5 were left for 0 days, 7 days, 14 days, 30 days and 60 days, appropriate amounts of samples were taken to prepare samples to be tested.
[0082] Take appropriate amount of the above samples, add 30mL 0.1mol / L sodium hydroxide solution respectively, dissolve by ultrasonic for 20min; dilute to 100mL with 0.1mol / L sodium hydroxide solution, centrifuge at 8000r / min for 5min, filter with 0.45μm filter membrane, collect the filtrate, which is the sample to be tested. For samples with a concentration higher than 10.5μg / mL, dilute to 2.1-10.5μg / mL with 0.1mol / L sodium hydroxide solution.
[0083] 5. Test the sample
[0084] The above-mentioned samples to be tested were injected according to the liquid chromatography test conditions, and the peak area of each sample to be tested was obtained from the liquid chromatogram. All tests were repeated three times and the average value of the results was taken.
[0085] 6. Data Analysis
[0086] According to the peak area of each sample to be tested, the concentration corresponding to each sample to be tested was obtained using the standard curve, and according to the dilution ratio, the actual content of hemoglobin in the products prepared in Examples 1-7 and Comparative Examples 1-5 after being placed for 0 days, 7 days, 14 days, 30 days and 60 days was obtained respectively.
[0087] Hemoglobin detection ratio (%) = actual hemoglobin content / theoretical hemoglobin content × 100%.
[0088] The higher the hemoglobin detection ratio, the smaller the proportion of hemoglobin that is oxidized and decomposed, and the more stable the hemoglobin.
[0089] 7. Test results
[0090] The test results are shown in Table 1 below.
[0091] Table 1 Test results
[0092]
[0093] From the results in Table 1, it can be seen that after the products prepared in Examples 1-5 were placed for 60 days, the heme detection ratio was still above 97.5%, indicating that the proportion of ferrous iron in heme oxidized to high-valent iron was small and heme was relatively stable; thus, it was shown that by adding an appropriate amount of superoxide dismutase and catalase to heme, the heme iron in heme can be effectively prevented from being oxidized and the stability of heme can be improved.
[0094] It can be seen from the results after 7 days, 14 days, 30 days and 60 days that the stability of the product prepared in Example 6-7 is worse than that of the product obtained in Example 5, indicating that superoxide dismutase and catalase can only effectively improve the stability of hemoglobin when they exist at the same time in an appropriate ratio, and too high or too low is not conducive to improving the stability.
[0095] After 30 days of storage, the hemoglobin detection ratio of the products prepared in Comparative Examples 1-5 has dropped to below 97.0%. After 60 days of storage, the hemoglobin detection ratio has further dropped significantly, all below 94.0%, indicating that a large proportion of the ferrous iron in the hemoglobin is oxidized to high-valent iron and the hemoglobin stability is poor.
[0096] It can be seen from the results after 60 days of storage that the stability of the products prepared in Comparative Examples 1-3 is slightly better than that of the products obtained in Comparative Examples 4-5, but significantly worse than that of the product obtained in Example 5, indicating that the addition of superoxide dismutase or catalase alone can only slightly improve the stability of heme, and superoxide dismutase and catalase must be added simultaneously in an appropriate proportion to significantly and effectively improve the stability of heme.
[0097] The above results indicate that the heme composition and pharmaceutical composition provided by the present application have high heme stability.
[0098] Each embodiment in this specification is described in a related manner, and the same or similar parts between the embodiments can be referenced to each other, and each embodiment focuses on the differences from other embodiments.
[0099] The above description is only a preferred embodiment of the present application and is not intended to limit the protection scope of the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application are included in the protection scope of the present application.
Claims
1. A heme composition, characterized in that include: Heme, superoxide dismutase and catalase; wherein the ratio of the mass of the heme, the enzyme activity of the superoxide dismutase and the enzyme activity of the catalase is 5g:3000-6000IU:1000-3000IU.
2. Use of the heme composition according to claim 1 in the preparation of a medicament for preventing or treating iron deficiency anemia.
3. A pharmaceutical composition, characterized in that It comprises the heme composition according to claim 1 and a pharmaceutically acceptable carrier.
4. The pharmaceutical composition according to claim 3, characterized in that The dosage form is oral dosage form.
5. The pharmaceutical composition according to claim 4, characterized in that The oral dosage form is selected from one of oral liquid, syrup, granules, capsules, tablets, powders, pills and dripping pills.
6. Use of the pharmaceutical composition according to claim 3 in the preparation of a medicament for preventing or treating iron deficiency anemia.
Citation Information
Patent Citations
Polyfunctional polyhemoglobin-enzyme complex
RU2432172C1
Methods for the synthesis of a modified hemoglobin solution
US20020099175A1