Pharmaceutical composition for preventing or treating hearing impairment containing cilostazol and ginkgo biloba extract as active ingredients
A technology for cilostazol and hearing impairment, applied in the field of pharmaceutical compositions for preventing or treating hearing impairment, to achieve the effect of treating noise-induced hearing impairment or ototoxic hearing impairment and inhibiting apoptosis
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Embodiment 1
[0055] Embodiment 1: According to the effect analysis of compound preparation of the present invention on hearing function and vestibular function
[0056] Preparation of compound preparation
[0057] Cilostazol (CS, SK Chemicals) and Ginkgo biloba extract (GbE, SK Chemicals) were mixed in a weight ratio of 5:4 and dissolved in 0.5% (w / v) carboxymethylcellulose sodium (CMCNa, Sigma) in an aqueous solution to prepare a composite preparation (hereinafter referred to as renexin) at a concentration of 18 mg / mL.
[0058] Application of sample
[0059] 52 SD (Sprague-Dawley) rats (200-250 g) aged 8-10 weeks were divided into the following 6 groups, and each drug was administered as follows.
[0060] -The first group: normal control group (n=3);
[0061] -The second group: cisplatin (CDDP) administration group alone (n=13);
[0062] -The third group: ranashin+CDDP administration group (n=12);
[0063] - The fourth group: Ginkgo biloba extract+CDDP administration group (n=12); ...
Embodiment 2
[0080] Example 2: Morphological evaluation of auditory and vestibular hair cells administered according to the compound formulation of the present invention
[0081] Morphological evaluation of the middle and bottom
[0082] The rats in each group of Example were anesthetized, and then their temporal bones were extracted. Under a dissecting microscope, in the cochlea The round window (round window) and oval window (oval window) were drilled, and the perilymphatic space (perilymphatic space) was perfused with 4% glutaraldehyde (EMgrade, place of origin, Inc.CA, USA) with a Pasteur pipette. Each sample was then fixed in 4% glutaraldehyde solution for 24 hours. The surface of the cochlea and the perilymphatic space were washed three times with phosphate buffered solution (PBS), perfused with 1% osmium tetroxide, and placed on a tissue rotator for 15 minutes. Then, the sample was washed 3 times with PBS, the bony capsule of the cochlea was removed under a dissecting microsco...
Embodiment 3
[0094] Embodiment 3: Analysis of the in vitro effect of the compound preparation of the present invention on CDDP-induced cytotoxicity
[0095] Confirmation of CDDP-induced cytotoxicity
[0096] In order to confirm CDDP-induced cytotoxicity, the auditory cell line (HEI-OC1 (ATCC, USA) was cultured at 33°C with 10% CO2 in Dulbecco's essential medium (DMEM, Dulbecco's minimum essential medium) was cultured. The cultured cells were divided into 96-well plates at a concentration of 3000 cells / well, and then 20 μM cisplatin was added for culture. At 0, 12, 24 and 48 hours of culture , add 20 μl of MTS solution to each well, continue to incubate for 4 hours, and then measure the absorbance of formazan at 490 nm with a fully automatic quantitative drawing microplate reader, so as to determine the proportion of surviving cells.
[0097]The measurement results are shown in Figure 8 . Such as Figure 8 As shown, according to the treatment with cisplatin, a time-dependent decrease ...
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