| Objective:Cholesterol-galactoside ligand is a newly synthesized galactoside ligand for liposomes which prepared by our laboratory.Now it has been put into liposome successfully.But the synthesis of cholesterol-galactoside ligand still remains in laboratory,it leads to a fact that the technology line is not perfect.The purpose of this article is to accomplish amplification for cholesterol-galactoside ligand,and investigate its corresponding paclitaxel liposome in acute toxicity study and stability study.Method:1.The step-by-step amplifying was used in the amplification of CHS-DD-LA,in this experiment,productivity and purity are the criteria.First,synthesis was enlarged to 5 times larger,and single factor study was used to find the best condition that productive>90%,purity>95%.Then using the same method in the study of 50 times enlarging.2.HPLC determination was selected for the content of paclitaxel in GAL-PTX-LP。Taxol(standard)was used as external standard.3.Transfering GAL-PTX-LP from liquid into lyophilized powder,particle size,potential,encapsulation efficiency and other indicators were used as criteria,then using method and condition that Chemical stability principle regulated to test its stability in storage;4.Transfering GAL-PTX-LP from liquid into lyophilized powder,the particle size,potential,encapsulation efficiencyand other indicators were used as criteria,tested its compatible stability with 0.9%Sodium Chloride Injection、5%Glucose Injection、10%Sodium Chloride Injection、50%Glucose Injection,time was set to be 24h,taking sample in 0、4、8、12、24h.5.GAL-PTX-LP lyophilized powder and ordinary paclitaxel liposome lyophilized powder were injected into rats(SPF)separately,0.9%Sodium Chloride Injection was chosedas solvent,single dose injection.It can be indicated in trial test about the safe clinical dose range.Then in official test,injected dose can be figured out with the safe clinical dose range by the formula below:(r:group rate,n:sections,a:minimal lethal dose,MLD,b:Maximal tolerance dose,MTD)Observing for 14 days after injection,and recorded death number of every group,calculated LD50 for two preparations.Results:1.In the 50 larger amplifying,the formulation and technology were determined through orthogonal test and selecting molar ratio of LA and cholesteryl hemi-vinyl sebacate,kind and amount of enzyme,ratio of solvent and other factors.The productive was about 96%,the purity was about 95%.2.The HPLC methodology study showed that impurities made no difference,the precision,recovery ratealso met tequirments.3.The stability test showed that the particle size,potential,encapsulation efficiency and other indicators of GAL-PTX-LP didn’t change too much in influencing factor experiment and in the accelerated stability experiment,these indicators were stable in 3 months.4.After dissolving in 0.9%Sodium Chloride Injection、5%Glucose Injection、10%Sodium Chloride Inject ion、50%Glucose Injection separately,the particle size,potential,encapsulation efficiency and other indicators of GAL-PTX-LP didn’t change too much in 24h.5.Acute toxicityexperiment showed that MLD and MTD of paclitaxel liposome was 30mg/mland 20mg/ml,the MLD and MTD of GAL-PTX-LP was 80mg/mland 40mg/ml.The LD50 of paclitaxel liposome was 23.454mg/ml,95%credibility interval was 22.109-24.788 mg/ml;the LD50 of GAL-PTX-LP was 63.455 mg/ml,95%credibility interval was 57.544-71.872 mg/ml.Conclusion:1.Cholesterol-galactoside ligand can be amplified to 50 larger productively and purely,so it can be applied to industrial production.2.HPLC determination can be used to detect content of paclitaxel in GAL-PTX-LP.3.The lyophilized powder GAL-PTX-LP is stable in extreme environment in short time,and it should not be storage longer than 3 months.4.Compatible stability experiment shows that GAL-PTX-LP is a good injection.5.Acute toxicity experiment shows that GAL-PTX-LP is safer than ordinary paclitaxel liposome when applied to clinic.It may be revalant to the hepatic targeting tendency of liposome. |
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