Preparation Of Thermosensitive In Situ Gel Of Glucosamine Hydrochloride For Intra-articular Injection And Its Pharmacological Effects On Osteoarthritis

Glucosamine is one of the commonly used drugs for the treatment of osteoarthritis.It has the functions of promoting proteoglycan synthesis,repairing damaged cartilage tissue,and resisting inflammation.At present,most of the preparations on the market are tablets and capsules of hydrochloride and sulfate,and the amount of the cartilage reaching the cartilage after oral administration is limited,which often affects the clinical effect.As a new drug delivery system,temperature-sensitive gel has the advantages of controlled release or sustained release,prolonged drug action time,localization targeting,etc.,and can be used for multiple routes of administration.This study designed a glucosamine hydrochloride thermosensitive gel that can be used for intra-articular injection,and studied its preparation method and related quality evaluation.At the same time,rabbit osteoarthritis was used as an experimental animal model to study the glucosamine hydrochloride temperature sensitive gel.The therapeutic effect of intra-articular injection on experimental osteoarthritis has achieved the intended research goal.1.To investigate the related properties of glucosamine hydrochloride.Different concentrations of glucosamine hydrochloride solution were prepared and their pH values were measured.The solution traits and drug content were used as indicators to investigate the effects of pH,low temperature and NaHSO₃ on the stability of glucosamine.The results showed that glucosamine hydrochloride was acidic?pH<5?after being dissolved in water.The pH of the solution was raised to 5,5.5 and 6.The solution turned yellow after being placed at room temperature for 10 days,and the solution was only yellowed at a low temperature?4??for 10 days.After the pH value was increased,the solution to which NaHSO₃ was added was left at room temperature for 10 days,and the properties were unchanged,and all of them were colorless and transparent.It shows that increasing the pH reduces the stability of glucosamine,while low temperature and NaHSO₃ can enhance its stability.2.The preparation process of glucosamine hydrochloride temperature sensitive gel was studied.The glucosamine hydrochloride temperature-sensitive gel was prepared by cold-dissolving method.The gelation temperature was used as the index to investigate the effect of poloxamer 407?P407?,poloxamer 188?P188?,glucosamine hydrochloride?GH?,NaHSO₃ and NaHCO₃ on temperature sensitive gel,and optimize the prescription with orthogonal experiments.The final formulation of the glucosamine hydrochloride temperature sensitive gel is:23%P407+6%P188+5%GH+0.2%NaHSO3,and NaHCO₃ adjusts pH=6.3.The quality of glucosamine temperature-sensitive gel was evaluated by appearance,gelation temperature,content,viscosity,needle penetration and in vitro release,and the stability was investigated by influencing factors.The results show that the temperature-sensitive gel prepared by the experiment was a colorless and transparent liquid at room temperature,but it was a colorless and transparent colloid at pHysiological temperature.The average gelation temperature is 35.4±0.094°C,and the average content of glucosamine is 47.96±0.26mg/mL.The temperature-sensitive gel with good needle penetration can be used for injection administration.In vitro release experiments showed that the cumulative release of aqueous glucosamine solution reached 100%at 3 h,while the glucosamine temperature-sensitive gel was completely released at 12 h,so the temperature-sensitive gel had a sustained release effect.The influencing factors experimental results show that under high temperature and strong illumination,the gelation temperature of the temperature sensitive gel increases,and the content decreases,but there is no obvious change under the condition of 4°C,suggesting that the prepared temperature sensitive gel should be sealed,low temperature and protected from light.4.The therapeutic effect of glucosamine hydrochloride thermosensitive gel on experimental osteoarthritis was investigated.Five of the 35 rabbits were selected as the normal physiological group,and no treatment was performed.The remaining 30rabbits were injected with papain in the knee joint cavity for osteoarthritis modeling.The modeled rabbits were divided into a high-dose gel group,a medium-dose gel group,a low-dose gel group,a glucosamine hydrochloride aqueous solution group,a blank gel group,and a model group.The intra-articular injection was administered once a week for 4 weeks.The width of the knee joint,the content of IL-1?,TNF-?,PG E2,NO in the joint fluid and the glycosaminoglycan content in the cartilage,and the changes of cartilage histology were selected as indicators.The results showed that papain successfully produced an osteoarthritis model:the knee joint was obviously swollen,the inflammatory factor content in the joint fluid was increased,and the cartilage structure was destroyed.At the end of the dosing period,compared with the model group,the knee joint width of the glucosamine-treated group decreased,the inflammatory factor content decreased significantly,the glycosaminoglycan and related proteoglycan content increased,and the cartilage structure was intact.Among the experimental groups,the effect of the medium dose gel group was better than that of the high dose group and the low dose group.The medium dose gel and the aqueous solution group were administered in the same amount,but the overall effect of the medium dose gel group was superior to that of the aqueous solution group.The above results indicate that glucosamine can inhibit the swelling of joints and the production of inflammatory factors in osteoarthritis,promote cartilage repair,and the gel effect is better than aqueous solution.In this study,the prescription of the temperature-sensitive gel containing glucosamine were successfully screened.The quality and pHarmacodynamics of temperature-sensitive gel was evaluated.It is simple to make the glucosamine temperature-sensitive gel.The thermosensitive gel has a good therapeutic effect on osteoarthritis,with the better effect than aqueous solution at the same dose.This study enriched the clinical dosage form of glucosamine as a veterinary drug,and provided a new idea for the development of veterinary osteoarthritis therapeutic drugs.