Application of cyanobacteria in treatment of kidney injury
By combining photosensitizers and other drugs with light-pretreated cyanobacteria, an injectable liquid formulation was prepared and directly injected into the kidneys, solving the treatment challenge of acute kidney injury and achieving safe and efficient kidney function recovery and cell protection.
Patent Information
- Application Number
- CN202511268490.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-05
- Publication Date
- 2025-10-17
AI Technical Summary
Existing technologies lack effective early intervention methods to treat kidney injury, especially acute kidney injury, which may progress to chronic kidney disease and ultimately lead to end-stage renal disease, causing great harm to patients' health and society.
Using light-pretreated cyanobacteria, combined with photosensitizers and other drugs for treating kidney injury, a liquid formulation is prepared for in situ injection to target acute kidney injury lesions. These drugs include ACEI/ARBs, diuretics, calcium channel blockers, glucocorticoids, immunosuppressants, antibiotics, analgesics, and traditional Chinese medicines, supplemented with pharmaceutically acceptable excipients. The formulation is an injectable liquid preparation that is directly injected into the damaged kidney site.
It enables targeted treatment of acute kidney injury, reduces the risk of side effects, provides a safer and more efficient treatment pathway, and significantly improves kidney function and reduces kidney cell apoptosis.
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Figure CN120789113A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the field of biological medicine, and particularly relates to application of cyanobacteria in treatment of kidney injury. BACKGROUND
[0002] Kidney injury is a kind of disease seriously affecting kidney function, including chronic kidney disease (CKD), acute kidney injury (AKI) and the like, which can cause proteinuria, edema and the like, seriously endangering health and needing timely diagnosis and treatment.
[0003] Among them, chronic kidney disease refers to kidney structure and function disorder caused by various reasons, kidney function is progressively decreased, mainly manifested as progressive irreversible loss of kidney units, reduced kidney regeneration capacity, metabolic changes, oxidative stress and inflammation, and finally leading to kidney fibrosis. Kidney fibrosis is the final result of various CKD and is one of main reasons causing end stage renal disease (ESRD). Acute kidney injury (AKI) is one of common critical diseases in clinic, and its diagnostic criteria mainly include serum creatinine (Scr) and urine volume indicators, and these indicators often appear in the progress period of disease, which may delay monitoring of early lesion process, and then make it progress to chronic kidney disease (CKD) and finally lead to end stage renal disease (ESRD), thereby causing great harm to health of patients themselves, family and society.
[0004] In recent years, the morbidity and mortality of the disease are continuously increasing, but its pathogenesis is complex, especially lacking effective early intervention means, which brings great difficulties to prevention and treatment of the disease. Therefore, it is of great significance to provide a new drug for treating kidney injury in the field. SUMMARY
[0005] In order to make up for the deficiency of the prior art, the application provides application of cyanobacteria in treatment of kidney injury.
[0006] To achieve the above object, the application adopts the following technical scheme:
[0007] The first aspect of the application provides application of cyanobacteria in preparation of a drug for treating kidney injury.
[0008] Further, the kidney injury is acute kidney injury.
[0009] Further, the cyanobacteria are cyanobacteria pre-processed by light.
[0010] Further, the drug further comprises a photosensitizer.
[0011] Further, the drug further comprises other drugs for treating kidney injury.
[0012] Further, the other drugs for treating kidney injury comprise ACEI / ARB drugs, diuretics, calcium channel blockers, glucocorticoids, immunosuppressants, antibiotics, analgesics, traditional Chinese medicines.
[0013] Further, the drug further comprises a pharmaceutically acceptable excipient.
[0014] Further, the drug is in a liquid dosage form.
[0015] Further, the liquid dosage form is a liquid injection preparation.
[0016] The second aspect of the present application provides a drug for treating kidney injury, the drug comprising cyanobacteria.
[0017] Further, the kidney injury is acute kidney injury.
[0018] Further, the cyanobacteria are cyanobacteria pre-treated by light.
[0019] The third aspect of the present application provides use of cyanobacteria in the preparation of a drug for regulating serum creatinine, beta2-MG, KIM-1, urea nitrogen.
[0020] Further, the cyanobacteria are cyanobacteria pre-treated by light.
[0021] The fourth aspect of the present application provides use of cyanobacteria in the preparation of a product for inhibiting kidney cell apoptosis.
[0022] Further, the cyanobacteria are cyanobacteria pre-treated by light.
[0023] Further, the inhibition of kidney cell apoptosis is non-therapeutic inhibition of kidney cell apoptosis in vitro.
[0024] The fifth aspect of the present application provides a method for treating kidney injury, the method comprising administering cyanobacteria.
[0025] Further, the kidney injury is acute kidney injury.
[0026] Further, the cyanobacteria are cyanobacteria pre-treated by light.
[0027] Advantages and beneficial effects of the present application:
[0028] The application breaks through the limitation that the existing cyanobacteria are only used for extracting metabolites, realizes that the living cyanobacteria target the lesions of acute kidney injury through in-situ injection, the method of the application has strong targeting and low risk of side effects, provides a new safe and efficient path for AKI treatment, and has a wide application prospect. BRIEF DESCRIPTION OF DRAWINGS
[0029] Figure 1 FIG. 1 is a characterization diagram of cyanobacteria, wherein 1A is a cyanobacteria morphology diagram under an optical microscope, 1B is a cyanobacteria morphology diagram under a fluorescence microscope, 1C is a cyanobacteria morphology diagram under a scanning electron microscope, and 1D is a cyanobacteria oxygen production diagram after light irradiation;
[0030] Figure 2 FIG. 2 is a cyanobacteria improved acute kidney injury renal function diagram, wherein 2A is a serum creatinine (CRE) content diagram, 2B is a β2-MG content diagram, 2C is a KIM-1 content diagram, 2D is a urea nitrogen (BUN) content diagram, 2E is a kidney paraffin section H&E staining diagram, and 2F is a kidney paraffin section TUNEL staining diagram. DETAILED DESCRIPTION
[0031] The following provides definitions of some terms used in the specification. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the application belongs.
[0032] The application provides application of cyanobacteria in preparation of a medicine for treating kidney injury.
[0033] In some embodiments, the kidney injury includes chronic kidney disease, acute kidney injury.
[0034] In a preferred embodiment, the kidney injury is acute kidney injury. The acute kidney injury is a disease in which the kidney function is sharply reduced between several hours to several days, and can be mainly divided into ischemic acute kidney injury and injury caused by nephrotoxic substances.
[0035] In some embodiments, the ischemic acute kidney injury is due to insufficient perfusion of blood flow to the kidney, leading to hypoxia and necrosis of renal tubular epithelial cells, and further triggering a clinical syndrome in which the kidney function is sharply reduced in a short period (several hours to several days). The ischemic acute kidney injury can be identified by a clinician, for example, by identifying ischemic conditions or by referring to certain inherent nephropathies of acute kidney injury.
[0036] In some embodiments, the core mechanism of the injury caused by nephrotoxic substances is that the nephrotoxic substances directly or indirectly damage the structure and function of the kidney. The nephrotoxic substances can include pesticides or drugs, contrast agents, etc.
[0037] In some embodiments, kidney diseases can be classified into prerenal, renal, and postrenal kidney diseases according to the site of injury. Prerenal kidney disease refers to kidney disease caused by reduced blood flow to the kidney due to systemic diseases, such as dehydration, shock, burns, massive hemorrhage, congestive heart failure, cirrhosis, renal artery stenosis, etc. Renal kidney disease refers to kidney disease caused by factors in the kidney itself, such as blood flow obstruction in the kidney, glomerular injury, and renal tubular / interstitial injury. Postrenal kidney disease refers to kidney disease caused by problems in the downstream urinary tract of the kidney.
[0038] In some embodiments, the cyanobacteria of the present application can be administered to the injury site after in vitro light pretreatment to treat kidney injury; or the cyanobacteria can be administered directly to the injury site and then subjected to light treatment to treat kidney injury; or the cyanobacteria can be engineered to treat kidney injury; or the cyanobacteria can be chemically pretreated to treat kidney injury.
[0039] In some embodiments, light pretreatment (or light pretreatment) is a technique for pretreating biological materials (such as the cyanobacteria of the present application) by light. The light used for light pretreatment can be any type of light. The types of light include, but are not limited to, sunlight, light sources simulating sunlight (such as fluorescent lamps, incandescent lamps, LED lamps), and monochromatic light of specific wavelengths (such as red light, blue light, ultraviolet light).
[0040] In some embodiments, light pretreatment can be continuous light, pulsed light, or gradient light. Continuous light refers to uninterrupted light, with constant light intensity and wavelength during the treatment. Pulsed light refers to irradiation in the form of intermittent light pulses (alternating light-dark), with constant light intensity during the pulse, but overall periodic changes. Gradient light refers to linear or nonlinear changes (such as stepwise enhancement, stepwise adjustment) in light intensity or wavelength over time, forming a dynamic light environment.
[0041] In some embodiments, the cyanobacteria are engineered, specifically, the cyanobacteria are genetically engineered with photosensitive proteins (such as bacterial rhodopsin), oxygen-producing enzymes (such as PSII complex in photosynthesis), or light-responsive elements (such as light-controlled promoters, light-sensitive transcription factors) introduced into the cyanobacterial genome; or photosensitizers (such as 5-ALA) are introduced into the cyanobacteria to prepare engineered cyanobacteria.
[0042] In some embodiments, chemical pretreatment refers to treating cyanobacteria with hydrogen peroxide (H2O2) or sodium percarbonate (Na2CO3·1.5H2O2).
[0043] In a preferred embodiment, the cyanobacteria of the present application are administered to the injury site after in vitro light pretreatment.
[0044] The drug also includes a photosensitizer.
[0045] In some embodiments, the photosensitizer includes but is not limited to 5-aminolevulinic acid, porphyrin derivatives.
[0046] The drug also includes other drugs for treating kidney injury.
[0047] The other drugs for treating kidney injury include but are not limited to ACEI / ARB drugs, diuretics, calcium channel blockers, glucocorticoids, immunosuppressants, antibiotics, analgesics, traditional Chinese medicines.
[0048] In some embodiments, the ACEI / ARB drugs include but are not limited to benazepril, fosinopril, captopril (ACEI), losartan, telmisartan, irbesartan (ARB).
[0049] The diuretics include but are not limited to furosemide, spironolactone, hydrochlorothiazide.
[0050] The calcium channel blockers include but are not limited to nifedipine, amlodipine, verapamil.
[0051] The glucocorticoids include but are not limited to prednisone, methylprednisolone, dexamethasone.
[0052] The immunosuppressants include but are not limited to cyclophosphamide, azathioprine, methotrexate.
[0053] The antibiotics include but are not limited to ceftriaxone, penicillin, levofloxacin.
[0054] The analgesics include but are not limited to tramadol (non-opioid central analgesics).
[0055] The traditional Chinese medicines include but are not limited to Bailin capsules, Jinshuibao capsules, Niaoduqing granules.
[0056] The drug also includes pharmaceutically acceptable excipients.
[0057] In some embodiments, the pharmaceutically acceptable excipients refer to those compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use with human and animal tissues without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.
[0058] Pharmaceutically acceptable excipients include saline, aqueous buffer solutions, solvents, and / or dispersion media. The use of such excipients is well known in the art, and solutions are preferably sterile fluids, and are generally in isotonic, physiologically compatible buffers. Such solutions are preferably fluid and transdermally administrable amounts, and are presented in unit or multi-dose sealed containers, for example, ampules or vials. The solutions are preferably stable under the conditions of manufacture and storage and preserved against the contaminating action of microorganisms, such as bacteria and fungi, by the use of preservatives, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like.
[0059] The pharmaceutical is in a liquid dosage form.
[0060] In preferred embodiments, the liquid dosage form is a liquid formulation for injection.
[0061] In some embodiments, the administration of the pharmaceutical of the present application is not particularly limited, for example, subcutaneous injection, intradermal injection, intramuscular injection, intralymph node injection, intravenous injection, intra-arterial injection, intraperitoneal injection, intrathoracic injection, direct local injection, and the like.
[0062] In preferred embodiments, the administration is direct local injection, which is direct injection into a target tissue / organ. In the present application, it is direct injection into a damaged kidney site.
[0063] In some embodiments, the injection can be injected by an injection pump, or can be delivered with the aid of an interventional catheter.
[0064] In some embodiments, the dosage of the pharmaceutical can vary from patient to patient. Determining the optimal dosage typically involves balancing the level of therapeutic benefit against any risk or deleterious side effects. The selected dosage level will depend on a variety of factors including, but not limited to, the activity of the pharmaceutical, the route of administration, the time of administration, the rate of excretion of the pharmaceutical, the duration of the treatment, other drugs, compounds, and / or materials used in combination, the severity of the disease, and the species, sex, age, weight, disease, general health, and prior medical history of the patient. The amount of the pharmaceutical and the route of administration are ultimately decided by a physician, veterinarian, or clinician, but are generally selected to achieve local concentrations at the site of action that are associated with desired effects without causing substantial harmful or deleterious side effects.
[0065] The administration can be continuous or intermittent (for example, at appropriate intervals to provide separate doses) throughout the course of treatment at one dosage. Methods of determining the most effective means and dosage of administration are well known to those of skill in the art and vary with the formulation, the purpose of the treatment, and the subject being treated. Administration can be one or more times at a dosage level chosen from the range of the physician, veterinarian, or clinician.
[0066] The application will be further described in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are presented by way of example and not as limitations to the present application. The principal features of the present application can be employed in various embodiments without departing from the scope of the present application.
[0067] Example 1 Characterization of cyanobacteria
[0068] 1. Experimental materials
[0069] The experimental materials used in this example and their sources are shown in Table 1.
[0070] Table Experimental materials used in Example 1
[0071]
[0072] 2. Experimental methods
[0073] 2.1 Cyanobacteria culture: The shaker temperature for cyanobacteria culture was 30°C, normal air environment culture, the shaker speed was 125 rpm / min, and the light in the shaker was turned on.
[0074] 2.2 Collection of cyanobacteria: 0.5 ml of cyanobacteria stock solution was taken and placed in an EP tube, centrifuged at room temperature at 8000 rpm / min for 5 min, and the cyanobacteria were precipitated at the bottom of the tube. The transparent supernatant was discarded, and PBS was added to resuspend and mix well. The supernatant was again centrifuged under the same conditions and discarded. This was repeated three times to wash away the trace elements in the cyanobacteria.
[0075] 2.3 Morphological observation of cyanobacteria: A small amount of cyanobacteria was taken using a pipette and evenly dropped on a glass slide, and a cover glass was used to press the slide. The slide was observed and photographed under a normal light microscope and a fluorescence microscope, respectively. Ultra-micro morphology: After the cyanobacteria were collected, they were subjected to gradient dehydration, staining and fixing, and other scanning electron microscope sample preparation steps. After the end of the preparation steps, the glass slide was cut into small pieces and observed under a scanning electron microscope to observe the ultra-micro morphology of the cyanobacteria.
[0076] 2.4 Dissolved oxygen detection of cyanobacteria: 1 x 10 6 cyanobacteria were resuspended in PBS. The HANSA Oxytrace+ oxygen electrode instrument was turned on and calibrated according to the steps. After the oxygen electrode calibration was completed, the resuspended cyanobacteria were added, and the dissolved oxygen content was detected. After a period of time, the cyanobacteria were irradiated with light (0.5 W / cm 2 ), and the dissolved oxygen content was continuously detected. The results showed that after receiving light, the cyanobacteria began to produce more dissolved oxygen, which could last for a period of time.
[0077] 3. Experimental results
[0078] Cyanobacteria appear green and long under an optical microscope ( Figure 1 A); Observation of cyanobacteria's spontaneous red fluorescence under a fluorescence microscope ( Figure 1 B); Scanning electron microscopy shows that a single cyanobacterium is in the shape of a slender rod ( Figure 1 C); Cyanobacteria can produce a large amount of dissolved oxygen after exposure to light ( Figure 1 D).
[0079] Example 2 Cyanobacteria improve renal function in mice with ischemic acute kidney injury
[0080] 1. Experimental Materials
[0081] The experimental materials and sources used in this example are shown in Table 2.
[0082] surface Experimental materials used in Example 2
[0083]
[0084] 2. Experimental Methods
[0085] Cyanobacteria were collected and counted in advance. At least 30 minutes before the injection of cyanobacteria into mice, the counted cyanobacteria were illuminated with LED light (light intensity 0.5 W / cm 2 ) for light pretreatment to ensure that the cyanobacteria were in an efficient oxygen-rich state before injection. The mice were randomly divided into three groups: NC (control), AKI (acute kidney injury), and AKI+S (acute kidney injury + cyanobacteria). Each group was treated as follows:
[0086] ① All groups first exposed the mouse kidneys through surgery;
[0087] ② In the NC group, only the kidney was exposed and 20 μL of normal saline was injected as the control group.
[0088] ③AKI group: The mouse kidneys were surgically exposed, and the bilateral renal arteries of the mice were clamped for 30 minutes to establish an AKI model. Under light conditions, 20 μL of normal saline was injected in situ into the mouse kidneys.
[0089] ④AKI+S group: The model was established according to the AKI group method. Under light conditions, pre-activated cyanobacteria (5.0×10 5 resuspended in 20 μL of saline).
[0090] After the operation, the mice were sutured and allowed to recover naturally. Twenty-four hours after surgery, all mice were sacrificed, and blood and kidney tissue samples were collected for subsequent experimental analysis.
[0091] The serum of the mouse blood was collected after centrifugation, and the serum was used for kidney function detection by using a kidney function detection kit; after the kidney tissue was made into a paraffin section, H&E staining and TUNEL immunohistochemistry were performed to evaluate the pathological changes and apoptosis of the mouse kidney tissue.
[0092] 3. Experimental results
[0093] The results of the kidney function detection of the mouse serum showed that the AKI mouse model was successful, the nitrogen metabolites (CRE and BUN) in the serum increased, the expression of the kidney injury related molecules increased, and the marker protein increased, while in the mice added with cyanobacteria, the serum nitrogen metabolites (creatinine and urea nitrogen) decreased, the kidney injury related molecules also decreased significantly, and the kidney function recovered to a certain extent. Figure 2 The H&E staining results of the mouse showed that the renal tubular lumen of the normal mouse kidney was small, the cell morphology was full, and the surface had cilia for reabsorption, the renal tubular lumen of the AKI mouse was large, the renal tubular epithelial cells appeared to fall off, the cell surface cilia disappeared, the cells were vacuolated, the cell nucleus was exposed and fell off, and the cell damage was obvious, while the above-mentioned phenomenon of the mouse added with cyanobacteria was alleviated. Figure 2 The TUNEL staining results showed that the apoptosis of the kidney cells of the AKI mouse increased, and the apoptosis of the kidney cells of the mouse added with cyanobacteria decreased. Figure 2 The histopathology results showed that the cyanobacteria alleviated the damage of the cells of the kidney tissue.
[0094] The above description of the embodiments is only for understanding the method of the present application and its core idea. It should be noted that for those skilled in the art, without departing from the principles of the present application, the present application can be improved and modified in several ways, and these improvements and modifications will also fall within the protection scope of the claims of the present application.
Claims
1. Application of cyanobacteria in the preparation of drugs for treating kidney damage; Preferably, the kidney injury is acute kidney injury.
2. The use according to claim 1, characterized in that The cyanobacteria are light-pretreated cyanobacteria.
3. The use according to claim 1, characterized in that The drug also includes a photosensitizer.
4. The use according to claim 1, characterized in that The drugs also include other drugs for treating kidney damage.
5. The use according to claim 4, characterized in that The other drugs for treating kidney damage include ACEI / ARB drugs, diuretics, calcium channel blockers, glucocorticoids, immunosuppressants, antibiotics, analgesics, and Chinese patent medicines.
6. The use according to claim 1, characterized in that The drug also includes pharmaceutically acceptable excipients.
7. The use according to claim 1, characterized in that The dosage form of the drug is a liquid dosage form; Preferably, the liquid dosage form is a liquid preparation for injection.
8. A drug for treating kidney damage, characterized in that: The drug includes cyanobacteria; Preferably, the kidney injury is acute kidney injury; Preferably, the cyanobacteria are light-pretreated cyanobacteria.
9. Application of cyanobacteria in the preparation of drugs for regulating serum creatinine, β2-MG, KIM-1, and urea nitrogen; Preferably, the cyanobacteria are light-pretreated cyanobacteria.
10. Application of cyanobacteria in the preparation of products for inhibiting renal cell apoptosis; Preferably, the cyanobacteria are light-pretreated cyanobacteria.