Use of glpg0187 in combination with propranolol for the preparation of a medicament for the treatment or inhibition of hemangiomas
By combining GLPG0187 and propranolol, the proliferation, migration, and tube formation of hemangioma endothelial cells are inhibited, solving the problem of side effects of propranolol monotherapy in infantile hemangiomas and achieving a safer and more effective treatment.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- NANTONG UNIV
- Filing Date
- 2026-03-17
- Publication Date
- 2026-06-09
Smart Images
Figure CN122163608A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of GLPG0187 in combination with propranolol in the preparation of drugs for the treatment or inhibition of hemangiomas. Background Technology
[0002] Infantile hemangiomas are the most common benign tumors in infancy, with an incidence rate of approximately 5% to 10%, and a male-to-female ratio of 1:3 to 1:5. Their development can be divided into three phases: proliferative, involution, and complete involution. The proliferative phase typically occurs within the first few months after birth, characterized by abnormal proliferation of vascular endothelial cells, leading to excessive angiogenesis. Most hemangiomas regress spontaneously within 1 to 5 years, eventually being replaced by fibrous adipose tissue, which can persist until age 12. Hemangiomas commonly occur in the head, neck, and limbs. Their growth rate varies from individual to individual; some grow slowly, while others can grow rapidly within weeks, leading to severe deformities. They are more common in women, premature infants, and low birth weight infants. Risk factors include advanced maternal age, placental abnormalities, multiple pregnancies, and exposure to certain medications or substances during pregnancy.
[0003] Currently, the oral beta-blocker propranolol is a first-line drug for treating infantile hemangiomas. However, approximately 2.1% of patients experience intolerable side effects after propranolol treatment, including sleep disturbances, agitation, bronchial hyperresponsiveness, bronchospasm, and hypoglycemia. Univariate regression analysis revealed that risk factors for ISEs include early age, prematurity, and low birth weight. Non-selective beta-blockers inhibit catecholamine-induced glycogenolysis, gluconeogenesis, and lipolysis, leading to hypoglycemia in the early stages of treatment. Furthermore, beta-blockers can also inhibit bronchial smooth muscle relaxation, causing respiratory side effects.
[0004] Integrin β1 (ITGB1) is an integrin located on human chromosome 10p11.22, encoding five isoforms. GLPG0187 is a small molecule inhibitor of integrin β1. Currently, there are no reports on the use of GLPG0187 in combination with propranolol in the treatment of hemangiomas. Summary of the Invention
[0005] In view of the problems or deficiencies in the existing technology, the present invention provides the application of GLPG0187 in combination with propranolol in the preparation of drugs for the treatment or inhibition of hemangiomas. Its application in anti-hemangioma drugs is expected to reduce the dosage of propranolol, provide a safer and more effective treatment for infantile hemangiomas, and improve patients' prognosis and quality of life.
[0006] To achieve the above-mentioned objectives, the technical solution adopted by this invention is as follows: The use of GLPG0187 in combination with propranolol in the preparation of drugs for the treatment or inhibition of hemangiomas.
[0007] Preferably, GLPG0187 is an integrin β1 inhibitor and propranolol is a β-receptor blocker.
[0008] Preferably, the effective concentration of GLPG0187 is 4 nM, and the effective concentration of propranolol is 20-30 μM.
[0009] Preferably, the drug exerts its therapeutic or inhibitory effect by inhibiting the proliferation, migration, or tube formation of hemangioma endothelial cells.
[0010] Preferably, the inhibition of hemangioma endothelial cell proliferation is verified by CCK8 assay; the inhibition of hemangioma endothelial cell migration is verified by scratch assay; and the inhibition of hemangioma endothelial cell tube formation is verified by tube formation assay.
[0011] A pharmaceutical composition for treating or inhibiting hemangiomas, comprising the above-mentioned GLPG0187 and propranolol.
[0012] Preferably, the dosage form of the pharmaceutical composition is an oral dosage form, an injection dosage form, a powder dosage form, an ointment dosage form, or a transdermal patch dosage form. Beneficial effects
[0013] Compared with existing technologies, the application of GLPG0187 in combination with propranolol in the preparation of drugs for treating or inhibiting hemangiomas, as demonstrated in this invention, through a combination drug strategy of integrin β1 inhibitor and β-receptor blocker, and verified by CCK8 assay, scratch assay, and tube formation assay, confirms that GLPG0187 combined with propranolol can further inhibit the proliferation, migration, and tube formation ability of human hemangioma endothelial cells, with better efficacy than propranolol monotherapy. This invention provides a new combination drug regimen for reducing the clinical dosage of propranolol and minimizing the risk of its side effects, offering a safer and more effective drug option for the treatment of infantile hemangiomas. Attached Figure Description
[0014] Figure 1 The image shows the CCK-8 proliferation experiment results after GLPG0187 was treated with propranolol for 24 hours. Figure 2 The image shows the scratch test results of HemECs treated with GLPG0187 in combination with propranolol; among them, Figure 2 A represents photomicrographs of the scratch width at 0 h, 12 h, and 24 h. Figure 2 B is a statistical chart of the 24-hour damage healing rate; Figure 3 The figure shows the tube formation results of HemECs treated with GLPG0187 in combination with propranolol; among them, Figure 3 A is a tube micrograph. Figure 3B is a statistical chart of the total length of the small tubes. Detailed Implementation
[0015] To make the technical problems, technical solutions and advantages of the present invention clearer, a detailed description will be given below in conjunction with specific embodiments.
[0016] Example 1, Experimental Materials Table 1 Cell Culture Reagents
[0017] Table 2 Reagents for Basic Experiments
[0018] Example 2, Experimental Method 1. CCK-8 Experiment (1) After digestion, the HemECs treated with the drug and the control group were counted and seeded into 96-well plates at 1600 cells / well and 3 replicates, and cultured for 24 hours.
[0019] (2) Prepare CCK-8 diluted with DMEM and add it to the 96-well plate. The final concentration of CCK-8 in each well is 10%. Add 3 blank wells separately to eliminate background error and continue incubation for 3-4 hours.
[0020] (3) Start the computer and microplate reader, put in a 96-well plate, and measure the absorbance at the specified wavelength (450nm).
[0021] 2. Scratch test (1) The drug-treated HemECs and the control group were plated in 6-well plates with 50% confluence and cultured in a 37℃, 5% CO2 incubator for 24h.
[0022] (2) Disinfect the ruler, observe under a microscope that the cell confluence reaches 80%, remove the culture plate, and use a 10 μL pipette to draw three lines per well, perpendicular to the bottom of the plate and close to the ruler. Add PBS to rinse and remove cell debris, remove waste liquid, and repeat rinsing once depending on the cell condition. Add DMEM, observe the scratches under an inverted phase contrast microscope, and take 4× photos to record the initial scratches for each group, taking three photos per well. After completion, return the plate to the cell culture incubator and let it stand. Take photos again at regular intervals afterward.
[0023] 3. Tube forming experiment (1) Prepare one day in advance: Melt the matrix adhesive in a 4℃ refrigerator, and pre-cool the ice box and pipette tip.
[0024] (2) Mix DMEM and matrix gel at a ratio of 1:1, add 50 μL / well to a 96-well plate, avoid generating air bubbles, place it flat in a 37°C incubator for at least 30 min, and wait for the matrix gel to solidify.
[0025] (3) After digestion and drug treatment, HemECs and control group were counted separately, and 26,000 cells / well were gently added to the substrate gel and cultured for 4-6 hours. During this period, the tube formation could be observed.
[0026] (4) Observe the tube formation of each well under a microscope at 4×, and take pictures for recording. Measure relevant parameters using ImageJ.
[0027] Example 3: Inhibitory effect of GLPG0187 combined with propranolol on the proliferation of human hemangioma endothelial cells HemECs in good growth condition were collected and, according to the CCK-8 assay method described in Example 2, the following treatment groups were set up: blank control group, propranolol monotherapy group (20 μM, 30 μM), GLPG0187 monotherapy group (4 nM), and GLPG0187 combined with propranolol group (4 nM GLPG0187 + 20 μM propranolol, 4 nM GLPG0187 + 30 μM propranolol). After 24 hours of treatment, the absorbance of each group of cells at 450 nm was measured to reflect the cell proliferation status.
[0028] Experimental results are as follows Figure 1 As shown in the figure, compared with the blank control group, the cell proliferation activity of the propranolol monotherapy group decreased in a concentration-dependent manner; the 4 nM GLPG0187 monotherapy group also showed a certain inhibitory effect on proliferation. Compared with the corresponding concentrations of propranolol monotherapy, the inhibitory effect of 4 nM GLPG0187 combined with 20 μM or 30 μM propranolol on HemEC proliferation was further enhanced. The results indicate that GLPG0187 combined with propranolol can synergistically inhibit the proliferation of human hemangioma endothelial cells.
[0029] Example 4: Inhibitory effect of GLPG0187 combined with propranolol on the migration of human hemangioma endothelial cells Following the scratch test method described in Example 2, HemECs were subjected to the same grouping treatment as in Example 3. The scratch healing status was observed and photographed at 0 h, 12 h, and 24 h after scratching, and the 24-hour damage healing rate was calculated.
[0030] Experimental results are as follows Figure 2 As shown. Figure 2 A shows photomicrographs of the scratch width at 0 h, 12 h, and 24 h for each treatment group. Figure 2B represents the statistical results of the 24-hour injury healing rate. Compared with the blank control group, the cell migration ability of the propranolol monotherapy group decreased in a concentration-dependent manner; the 4 nM GLPG0187 monotherapy group also showed a certain degree of migration inhibition. Compared with the corresponding concentrations of propranolol monotherapy, the inhibitory effect of 4 nM GLPG0187 combined with 20 μM or 30 μM propranolol on HemEC migration was further enhanced. The results indicate that GLPG0187 combined with propranolol can synergistically inhibit the migration of human hemangioma endothelial cells.
[0031] Example 5: Inhibitory effect of GLPG0187 combined with propranolol on the tube-forming ability of human hemangioma endothelial cells Following the tube formation experiment method described in Example 2, HemECs were grouped and treated in the same way as in Example 3, then inoculated onto Matrigel substrate. After culturing for 4-6 hours, the tube formation was observed, and the total tube length was counted using ImageJ software.
[0032] Experimental results are as follows Figure 3 As shown. Figure 3 A shows the tube micrographs of each treatment component. Figure 3 B represents the statistical results of total tubule length. Compared with the blank control group, the tubular formation ability of cells in the propranolol monotherapy group decreased in a concentration-dependent manner; the 4 nM GLPG0187 monotherapy group also showed a certain inhibitory effect on tubular formation. Compared with the corresponding concentrations of propranolol monotherapy, the inhibitory effect of 4 nM GLPG0187 combined with 20 μM or 30 μM propranolol on HemEC tubular formation was further enhanced. The results indicate that GLPG0187 combined with propranolol can synergistically inhibit the tubular formation ability of human hemangioma endothelial cells.
[0033] The above description represents the preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.
Claims
1. Application of GLPG0187 in combination with propranolol in the preparation of drugs for the treatment or inhibition of hemangiomas.
2. The application according to claim 1, characterized in that, GLPG0187 is an integrin β1 inhibitor, and propranolol is a β-receptor blocker.
3. The application according to claim 1, characterized in that, The effective concentration of GLPG0187 is 4 nM, and the effective concentration of propranolol is 20-30 μM.
4. The application according to claim 1, characterized in that, The drug exerts its therapeutic or inhibitory effects by inhibiting the proliferation, migration, or tube formation of hemangioma endothelial cells.
5. The application according to claim 1, characterized in that, The inhibition of hemangioma endothelial cell proliferation was verified by the CCK8 assay; the inhibition of hemangioma endothelial cell migration was verified by the scratch assay; and the inhibition of hemangioma endothelial cell tube formation was verified by the tube formation assay.
6. A pharmaceutical composition for treating or inhibiting hemangiomas, characterized in that, The pharmaceutical composition comprises GLPG0187 and propranolol as described in claim 1.
7. A pharmaceutical composition for treating or inhibiting hemangiomas according to claim 6, characterized in that, The dosage form of the pharmaceutical composition is oral, injectable, powder, ointment, or transdermal patch.