In Vitro Study Of Anti-osteoporosis Drugs Made Of Hyaluronic Acid-polylactic Acid Nanoparticles As Candidate Drugs

Microbial metabolites are a treasure trove for finding small-molecule drugs.Compared with plants and microorganisms,they can reproduce rapidly and meet the requirements of mass production of effective compounds.Japanese scholars discovered a compound Stigmastane-3-6-dione with anti-osteoclast activity against Streptomyces oxysporum with molecular formula C₂₉H₂₈O₂,which is resistant to the osteoclast differentiation rate,and its efficacy reached 49%.Shandong University scholars from China A steroidal compound isolated from the powder fungus,which is identical to the structure of the compound analyzed by Japanese scholars,but no activity studies have been conducted.The laboratory hopes to isolate the metabolizing compound from Ustilago maydis and coat it with Hyaluronic Acid-polylactic Acid Nanoparticle,using a certain molecular weight of hyaluronic acid against osteoclast active knots and small molecule compounds for the basic research of osteoporosis treatment.（1）Fermentation of corn smut by shaking flask,centrifuge the broth at 6000rpm/min for 5 min,take the supernatant and add 1.2 times of ethyl acetate,extract three times,take the organic layer,and evaporate the organic layer under reduced pressure.The column was chromatographed on a 1 cm silica gel column.The mobile phase was petroleum ether:ethyl acetate.The gradients were set to 1:0,6:1,2:1,and0:1,respectively.The 12 components are named Fr1-Fr12.The Fr2,Fr5,Fr6,and Fr7were separated using HPLC and RP-18 columns,and the main components were collected and designated as 2-0,2-1,2-2,5-0,5-1,and 5-2.5-3,6-0,6-1,6-2,7-0,7-1,identified by a primary mass spectrometry and secondary mass spectrometry,the 7-0sample is a Stigmastane-3-6-dione.（2）The collected samples were tested for anti-osteoclast activity and 7-0samples were found to have good anti-osteoclast differentiation activity.（3）Acid hydrolysis was used to hydrolyze hyaluronic acid to 1.4×10⁵ Da and chemical synthesis was performed with polylactic acid.After stirring,nanoparticles were prepared and analyzed for particle size,Zeta potential,and electron microscopy to determine that they meet the requirements as a drug carrier.（4）Select Alendronate as a drug for comparison,use the synthesized HA-PLA nanoparticles to coat Alendronate and 7-0 sample for anti-osteoclast differentiation experiments.The experimental results show that HA-PLA nanoparticles coated 7-0The drug-loaded nanoparticles prepared from the samples have good anti-osteoclast differentiation effect,and the efficacy is similar to that of alendronate.In this experiment,the anti-osteoclast active compounds were successfully purified and coated into HA-PLA nanoparticles to study the anti-osteoclast differentiation.It was proved that the anti-osteoclast-responsive compound had the anti-osteoclast differentiation effect when used alone and with the drug carrier.Complete a preliminary study of anti-osteoporosis treatment.