CN115590896B - Application of low molecular dextran bezoar acid and chlorella in preparing medicine for treating skin related diseases - Google Patents
Application of low molecular dextran bezoar acid and chlorella in preparing medicine for treating skin related diseases Download PDFInfo
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- A61K36/05—Chlorophycota or chlorophyta (green algae), e.g. Chlorella
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
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- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/716—Glucans
- A61K31/721—Dextrans
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
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- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
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- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/70—Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
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Abstract
The invention relates to an application of low molecular dextran bezoar acid and chlorella to the combined preparation of a medicament for treating skin related diseases. Wherein the skin related diseases involved include: burn and scald, pressure sore, diabetes ulcer, and atopic dermatitis; wherein, firstly, the low molecular dextran taurine solution is used for wetting wounds or skin, and then chlorella is used; wherein the low molecular dextran comprises dextran 20, 40, 60 and the like, and the dosage of taurine is 0.01-5% w/v; wherein the chlorella is applied in the forms of cells, preparations and the like, and specifically comprises a solution, hydrogel, patches, dressing and the like which take the chlorella as an active component; the application concentration of Chlorella is 1×10 3 ~1×10 12 cell/mL。
Description
Technical Field
The invention relates to application of chlorella, in particular to application of low-molecular dextran bezoar acid and chlorella in combined preparation of a medicament for treating skin related diseases, and in particular relates to burn and scald, pressure sore, diabetic ulcer and atopic dermatitis.
Background
Chlorella (Chlorella) its preparation methodChlorellasp.) is a single-cell eukaryotic algae (2-10 mu m), has extremely high growth speed, can resist various inhibitory growth conditions, and is one of the most widely cultivated microalgae species. Chlorella contains abundant nutrients such as protein, lipid, polysaccharide, amino acid, vitamins, etc., the protein content is up to 50%, and Chlorella has been developed as a human nutrition supplement. In addition, since the contents thereof further contain growth factors and immunomodulators, the composition can be used as health care products and functional foods for preventing diseases. Chlorella can continuously generate oxygen through photosynthesis under illumination conditions, and is recognized as a source of oxygen in the air. In addition, chlorella also has anti-inflammatory and antioxidant functions. The interior of the medicine contains abundant sulfated polysaccharide, and can play an anti-inflammatory role by activating and regulating macrophages. Besides sulfated polysaccharides, pigment-protein complexes in Chlorella extract also have anti-inflammatory effects.Currently, the identified inclusions with antioxidant and free radical scavenging functions include: lutein (lutein),𝛼-carotene,𝛽Carotene, ascorbic acid and𝛼tocopherol, etc. Furthermore, biologically active compounds thereof𝛽1,3 glucan as an active immunostimulant, which reduces cholesterol in blood, has also been shown to have a prophylactic function on atherosclerosis and hypercholesterolemia.
Chlorella has the ability to improve the microenvironment of the poor tissue of skin-related diseases, but is a living cell, and has high proliferation and metabolic activity, which may further consume nutrition of the lesion, reduce the nutrient supply of the tissue cells of the lesion, and further hinder the treatment of diseases. The low molecular dextran is a nutritional blood volume expanding medicine, and is suitable for treating diseases with hypovolemia and poor microcirculation due to protein deficiency. Wherein, the low molecular dextran can increase the osmotic pressure of plasma colloid, expand blood volume and expand capillary vessels, thereby improving blood perfusion and microcirculation of wound tissues, improving blood flow and increasing blood volume, and accelerating metabolism and discharge of harmful products thereof.
Taurine is an amino acid essential for human body conditions, and has been widely used in the fields of medicine, food additives, etc. Experiments at present prove that 0.01% -1% concentration taurine can promote fibroblast proliferation. In addition, taurine has anti-inflammatory and antioxidant effects, and can reduce the formation of peroxidized lipid, carbonyl protein and active oxygen by improving antioxidant enzyme activities such as superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and the like.
Therefore, we propose a therapeutic method using low molecular dextran taurine in combination with chlorella to obtain better therapeutic effect.
Disclosure of Invention
The invention relates to application of low-molecular dextran taurine and chlorella in combination in preparing a medicament for promoting wound healing.
The application of low molecular weight dextran taurine and chlorella in preparing medicine for treating skin diseases includes burn, scald, pressure sore, diabetes ulcer and atopic dermatitis.
The combined treatment method is to firstly apply low molecular dextran taurine and then apply chlorella.
The low molecular dextran taurine solution is characterized in that: every 1000, mL contains 60.0, g low molecular weight dextran 20, 40 or 60, the auxiliary material is sodium bisulphite, and the taurine content is 0.01-5% w/v.
The chlorella preparation comprises a solution taking chlorella as an active component, hydrogel, a patch, dressing, freeze-dried powder and the like.
The screening range of the chlorella biomass is 1 multiplied by 10 3 ~ 1 × 10 12 cell/mL。
The preparation method of the low molecular dextran taurine solution is characterized by comprising the following steps of: dissolving the low molecular dextran and taurine in normal saline, and sterilizing.
The chlorella solution comprises but is not limited to chlorella BG-11 solution, and the preparation method is characterized in that: centrifuging the sterile cultured chlorella with the determined biomass, discarding the supernatant, flushing the BG-11 culture medium for 2 times, and re-suspending by using the quantitative BG-11 culture medium to obtain the chlorella BG-11 solution.
The chlorella hydrogel comprises but is not limited to chlorella hyaluronic acid hydrogel, and the preparation method is characterized in that: 10 ml of 1X 10 8 Sterile cultured Chlorella at cell/ml concentration was centrifuged, the supernatant was discarded, BG-11 medium was washed 2 times, and then resuspended with a fixed amount of BG-11 medium. Then, 0.2g of hyaluronic acid powder was added to the chlorella solution, and stirred using a magnetic stirrer at room temperature to form chlorella hyaluronic acid hydrogel.
The combination of the low molecular dextran taurine and the chlorella is superior to the single application in the treatment effect.
The combination of the low-molecular dextran taurine and the chlorella is characterized in that the treatment effect of the combination can be remarkably improved by applying the low-molecular dextran taurine and then applying the chlorella.
Burns and scalds generally refer to the damage of tissues caused by heat, mainly skin and/or mucous membrane, and severe cases can also damage subcutaneous or/and submucosal tissues such as muscles, bones, joints and even viscera. The body is severely oxidized and damaged after burn and scald due to ischemia reperfusion injury of tissues and massive loss of body fluid of the body, so that a large amount of active oxygen metabolites are generated, and a severe inflammatory reaction is accompanied. In addition, burns and scalds can cause extensive physiological barrier damage of the organism, a large amount of tissue necrosis and reduced immune function, and meanwhile, a large amount of protein-rich exudates exist on the burn wound surface, so that the probability of invasion of the organism by pathogens is improved. The low molecular dextran and taurine can provide nutrition for wound tissues required by regeneration, the chlorella can obviously reduce oxidative stress and inflammation, has the function of purifying exudates by using metabolic proteins, and can also provide growth factors required by wound healing, so that the combination of the low molecular dextran and taurine can be a potential treatment strategy.
Pressure sores are ulcerated and necrotized tissues caused by continuous blood circulation blockage, hypoxia and nutrient deficiency due to long-term compression of local tissues, and are also called pressure ulcers and pressure sores. Skin pressure sores are a common problem faced in the long-term treatment process of rehabilitation treatment and nursing, and clinical treatment faces a certain difficulty, and about 6 ten thousand people die each year due to pressure sore combination symptoms. The low molecular dextran and taurine can provide nutrition for wound tissues for regeneration; the chlorella can be used for promoting revascularization, providing oxygen, providing nutrition, weakening sore face inflammation, and further promoting healing of pressure sores.
Diabetic ulcers are chronic wounds resulting from long-term hyperglycemia, with long treatment cycles and high recurrence rates, and patients are even at risk of amputation and death. Currently, patients worldwide are up to 2600 thousands of people, which causes a great medical burden. Studies have shown that diabetics account for 11.2% (about 1.298 billions) in our country adults, with the incidence of diabetic ulcers being about 4.1%. Diabetic chronic wounds are often manifested by high glucose, high oxidative stress, severe hypoxia, high lactic acid content and malnutrition. The low molecular dextran and taurine can provide nutrition for wound tissues required for regeneration and can improve insulin resistance and combine the characteristics of chlorella, so that the combination of the low molecular dextran and taurine seems to be a feasible method for improving the microenvironment of diabetic ulcer tissues.
Atopic dermatitis is a chronic recurrent inflammatory skin disease. Global disease burden studies showed that childhood prevalence is 15% to 20% and adult prevalence is as high as 10%, making atopic dermatitis the 15 th most common non-lethal disease, also the most economically burdened dermatological disease. It is usually manifested by itching, redness, swelling and dryness of the skin. Low molecular dextran and taurine can provide nutrition required for metabolism for inflammation parts, and chlorella can be used for providing oxygen and nutrients and a certain moist environment to accelerate the repair process of damaged skin, so that the combination of the low molecular dextran and taurine seems to be a feasible method for treating atopic dermatitis.
Advantageous effects
Low molecular dextran is commonly used for treating diseases with hypovolemia and poor microcirculation due to protein deficiency, and has limited effect although a certain therapeutic effect on wound healing, and does not show remarkable therapeutic advantages in the treatment of skin-related diseases. Taurine has anti-inflammatory and antioxidant effects, and can reduce insulin resistance of diabetic tissues. Chlorella has the ability to improve the microenvironment of the undesirable tissues of skin-related diseases, but also faces a certain dilemma.
The key points of the invention are as follows: the combination of the low-molecular dextran taurine and the chlorella can effectively treat burns and scalds, pressure sores, diabetic ulcers and atopic dermatitis, wherein the treatment effect of the combined application is obviously higher than that of the single application, and a synergistic effect is generated, and particularly the treatment effect is more obvious when the low-molecular dextran taurine is firstly applied and then the chlorella is applied.
1. The low molecular dextran taurine is combined with the chlorella, the active ingredients are cheap and easy to obtain, the preparation process is simple, and the method has industrial production potential.
2. The low molecular dextran taurine is combined with the chlorella, has remarkable treatment effect, and can effectively treat burns and scalds, pressure sores, diabetic ulcers and atopic dermatitis.
3. The combination of the low molecular dextran taurine and the chlorella has a treatment effect which is obviously higher than that of the single application of the low molecular dextran taurine or the chlorella.
4. In the combined application, the low-molecular dextran taurine is firstly applied and then the chlorella is applied, so that the compound has more remarkable treatment effect.
Drawings
FIG. 1 shows the application of the low molecular weight dextran taurine and the chlorella in the treatment of burns and scalds in combination in example 1 and example 2;
FIG. 2 shows the application of the low molecular weight dextran taurine and the chlorella in combination and the application of the low molecular weight dextran taurine and the chlorella in the treatment of pressure sores in example 1 and example 2;
FIG. 3 shows the combined application of low molecular weight dextran taurine and Chlorella, and the application of low molecular weight dextran taurine and Chlorella in the treatment of diabetic ulcer;
FIG. 4 shows the use of low molecular weight dextran taurine in combination with Chlorella, followed by low molecular weight dextran taurine in the treatment of atopic dermatitis according to example 1, example 2, and Chlorella.
Detailed Description
The present invention is further illustrated by the following specific examples, but the present invention is not limited to the following examples.
Example 1: preparation of low molecular weight dextran taurine solution
Adding normal saline (60.0 g dextran 40 and 1 g taurine in each 1000 mL normal saline) into a certain amount of low molecular weight dextran and taurine, stirring, and sterilizing to obtain low molecular weight dextran taurine solution.
Example 2: preparation of Chlorella hydrogel
10 ml of 1X 10 is taken 8 Sterile cultured Chlorella at cell/ml concentration, centrifuging (4000 rpm,3 min), discarding supernatantBG-11 medium was washed 2 times and resuspended in 10 ml BG-11 medium. Then, 0.2g of HA powder was added to the chlorella solution and stirred (500 rpm,4 h) at room temperature using a magnetic stirrer to form chlorella hyaluronic acid hydrogel.
Example 3: in vivo evaluation for treatment of burn and scald
A6-8 week old Balb/C male mouse is selected, and a constant temperature controlled voltage electric iron instrument is used for manufacturing a mouse scald model. After molding, the mice were randomly divided into five groups: burn and scald control group (wound sealed with 3M dressing), example 1 group (wound sealed with 3M dressing with 80 μl of low molecular weight dextran taurine solution), example 2 group (wound sealed with 3M dressing with 80 μl of chlorella hydrogel), combined application group (wound sealed with 3M dressing with 40 μl of low molecular weight dextran taurine solution, while wound sealed with 40 μl of chlorella hydrogel with 3M dressing), sequential application group (wound sealed with 3M dressing with 40 μl of low molecular weight dextran taurine solution first, wound sealed with 3M dressing with 40 μl of chlorella hydrogel after 1 h). All experimental groups changed hydrogels daily, the wounds were photographed on days 0, 3, 7, 10 and quantitatively analyzed by ImageJ software.
As shown in figure 1, the combined application of the low-molecular dextran taurine and the chlorella can effectively treat burns and scalds, and the combined treatment effect is obviously higher than that of the single application. Because the administration volume of the combined application group is unchanged (the total volume of the low-molecular dextran taurine and the chlorella is the same as that of the example 1 group and the example 2 group), but better treatment effect is obtained, the low-molecular dextran taurine and the chlorella are considered to have a certain synergistic effect in the combined treatment of burns and scalds. In addition, the application of the chlorella hydrogel after the application of the low-molecular dextran taurine has better treatment effect.
Example 4: in vivo evaluation of treatment of pressure sores
200 g SD male rats are selected, and a mechanical compression method is adopted to make a rat pressure sore model. After molding, the mice were randomly divided into five groups: pressure sore control (sealing wound with 3M dressing), example 1 (sealing wound with 80. Mu.L low molecular weight dextran taurine solution with 3M dressing), example 2 (sealing wound with 80. Mu.L Chlorella hydrogel with 3M dressing), combination application (sealing wound with 40. Mu.L low molecular weight dextran taurine solution with 40. Mu.L Chlorella hydrogel with 3M dressing), sequential application (sealing wound with 40. Mu.L low molecular weight dextran taurine solution first, 40. Mu.L Chlorella hydrogel after 1 h, sealing wound with 3M dressing). All experimental groups changed hydrogels daily, the wounds were photographed on days 0, 3, 7, 10 and quantitatively analyzed by ImageJ software.
As shown in figure 2, the combined application of the low-molecular dextran taurine and the chlorella can effectively treat pressure sores, and the combined treatment effect is obviously higher than that of the single application. Because the administration volume of the combined application group is unchanged (the total volume of the low-molecular dextran taurine and the chlorella is the same as that of the example 1 group and the example 2 group), but better treatment effect is obtained, the low-molecular dextran taurine and the chlorella are considered to have a certain synergistic effect when being combined for treating the pressure sore. In addition, the application of the chlorella hydrogel after the application of the low-molecular dextran taurine has better treatment effect.
Example 5: in vivo evaluation of treatment of diabetic ulcers
After 4-6 weeks old Balb/C male diabetic mouse feed (high fat and high sugar) was fed for 4 weeks, streptozotocin citrate buffer (streptozotocin: 55 mg/kg) was intraperitoneally injected for 5 consecutive days. Mice with post-detection blood glucose greater than or equal to 16.7. 16.7 mM were considered successful in modeling. Mice were anesthetized by intraperitoneal injection of chloral hydrate and shaved. A circular full-thickness skin wound was made using a biopsy punch of 8 mm inside diameter, and a silicone gasket was sewn around the wound tissue using non-absorbable sutures to prevent shrinkage of the mouse skin, hydrogel was added and a clear polyurethane film was selected as the outer backing layer to form a closed system. Mice were randomly divided into five groups in the experiment: the control group for diabetes (wound sealed with 3M dressing), the example 1 group (wound sealed with 3M dressing with 80. Mu.L of low molecular weight dextran taurine solution), the example 2 group (wound sealed with 3M dressing with 80. Mu.L of Chlorella hydrogel), the combination application group (wound sealed with 3M dressing with 40. Mu.L of low molecular weight dextran taurine solution, while wound sealed with 40. Mu.L of Chlorella hydrogel with 3M dressing), the sequential application group (wound sealed with 3M dressing with 40. Mu.L of low molecular weight dextran taurine solution first, 40. Mu.L of Chlorella hydrogel after 1 h). All experimental groups changed hydrogels daily. Wounds were photographed on days 0, 3, 7, 9 and quantitatively analyzed by ImageJ software.
As shown in figure 3, the combined application of the low-molecular dextran taurine and the chlorella can effectively treat the diabetic ulcer, and the combined treatment effect is obviously higher than that of the single application. Because the administration volume of the combined application group is unchanged (the total volume of the low-molecular dextran taurine and the chlorella is the same as that of the example 1 group and the example 2 group), but better treatment effect is obtained, the low-molecular dextran taurine and the chlorella are considered to have a certain synergistic effect in the combined treatment of the diabetic ulcer. In addition, the application of the chlorella hydrogel after the application of the low-molecular dextran taurine has better treatment effect.
Example 6: in vivo evaluation of treatment of atopic dermatitis
Base solutions were prepared with acetone and olive oil in a 3:1 ratio, and then 1% and 0.2% DNFB sensitizers were prepared with 2,4 Dinitrofluorobenzene (DNFB) and base solutions, with molding beginning for the first week and continuing for three weeks. After molding, the mice were randomly divided into five groups: the atopic dermatitis control group (wound sealed with 3M dressing), the example 1 group (wound sealed with 3M dressing with 80. Mu.L of low molecular weight dextran taurine solution), the example 2 group (wound sealed with 3M dressing with 80. Mu.L of chlorella hydrogel), the combination application group (wound sealed with 3M dressing with 40. Mu.L of low molecular weight dextran taurine solution, and 40. Mu.L of chlorella hydrogel with 3M dressing) and the sequential application group (wound sealed with 3M dressing with 40. Mu.L of low molecular weight dextran taurine solution first, 40. Mu.L of chlorella hydrogel after 1 h). The chlorella hydrogel was then used for treatment three times a week for a total of four weeks. After the experiment, the mice were sacrificed by cervical removal, and the ears and back skin were immersed in 4% paraformaldehyde fixing solution for pathological section.
The mice were scored for dermatitis at predetermined time points (0, 7, 14, 21, 25, 28) for back skin lesions. The specific criteria are as follows: asymptomatic, score 0; mild symptoms, 1 minute; moderate symptoms, 2 points; severe symptoms, 3 minutes. Erythema/bleeding, edema, scratch/erosion, and scaling/dryness symptoms were assessed separately, and dermatitis scores were calculated as the sum of individual scores, and were divided into 12 scores.
As shown in FIG. 4, the combined application of the low-molecular dextran taurine and the chlorella can effectively treat the atopic dermatitis, and the combined treatment effect is obviously higher than that of the single application. Since the administration volume of the combination application group is unchanged (the total volume of the low molecular weight dextran taurine and the chlorella is the same as that of the example 1 group and the example 2 group), but better treatment effect is obtained, the low molecular weight dextran taurine and the chlorella are considered to have a certain synergistic effect in combination treatment of atopic dermatitis. In addition, the application of the chlorella hydrogel after the application of the low-molecular dextran taurine has better treatment effect.
Claims (6)
1. The application of the combination of the low-molecular dextran taurine and the chlorella in preparing the medicine for promoting the wound healing is characterized in that the low-molecular dextran taurine is a low-molecular dextran taurine solution, and the solution is prepared by the following steps: every 1000 mL contains 60.0 g low molecular dextran 20, 40 or 60, the auxiliary material is sodium bisulphite, the taurine content is 0.01% -5% w/v, the low molecular dextran and taurine are dissolved in normal saline, and the sterilization is carried out; the chlorella is dressing made of chlorella.
2. The application of the combination of the low molecular dextran taurine and the chlorella in preparing the medicine for treating the skin diseases is characterized in that the skin diseases are burns and scalds, pressure sores, diabetic ulcers or atopic dermatitis; the low molecular weight dextran taurine is a low molecular weight dextran taurine solution, and the solution is prepared by the following steps: every 1000 mL contains 60.0 g low molecular dextran 20, 40 or 60, the auxiliary material is sodium bisulphite, the taurine content is 0.01% -5% w/v, the low molecular dextran and taurine are dissolved in normal saline, and the sterilization is carried out; the chlorella is dressing made of chlorella.
3. The use according to claim 1 or 2, wherein the dressing is a cell solution, a hydrogel or a patch.
4. The use according to claim 3, wherein the hydrogel is prepared from chlorella and hyaluronic acid.
5. The use according to claim 4, wherein the chlorella biomass screening range is 1X 10 3 ~ 1×10 12 cell/mL。
6. The use according to claim 5, wherein the hydrogel comprises chlorella hyaluronic acid hydrogel, the chlorella hyaluronic acid hydrogel being prepared by the steps of: 10 ml of 1X 10 8 Sterile cultured chlorella at the cell/ml concentration was centrifuged, the supernatant was discarded, BG-11 medium was washed 2 times, and re-suspended with a fixed amount of BG-11 medium, then, hyaluronic acid powder of 0.2. 0.2g was added to the chlorella solution, and stirred using a magnetic stirrer at room temperature to form chlorella hyaluronic acid hydrogel.
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| CN110859253A (en) * | 2019-10-10 | 2020-03-06 | 内蒙古阿尔格生命科学有限公司 | A beverage containing Chlorella and Stichopus japonicus extract |
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| US20100239655A1 (en) * | 2004-12-09 | 2010-09-23 | Georgia Levis | Taurine-based compositions and therapeutic methods |
| CN113924132B (en) * | 2019-05-07 | 2023-07-25 | 罗盖特公司 | Novel polysaccharide-based hydrogel scaffold for wound care |
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| CN1919350A (en) * | 2006-09-18 | 2007-02-28 | 俞锋 | New pattern compress for treating skin mucosa injury and its preparation method |
| JP2009091349A (en) * | 2007-09-19 | 2009-04-30 | Ogaki Bio Technology Kenkyu Center:Kk | External preparation for skin, bath preparation or cosmetics containing chlorella extract, and process for their preparation |
| CN110859253A (en) * | 2019-10-10 | 2020-03-06 | 内蒙古阿尔格生命科学有限公司 | A beverage containing Chlorella and Stichopus japonicus extract |
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| CN115590896A (en) | 2023-01-13 |
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