Studies On Toxicology Of Andrographalide Nanosuspension

The main component of andrographolide nano-suspension for injection is andrographolide,which is a new drug released system developed by a new process and technology.Its dosage form,excipients and production process are different from those of the drugs with andrographolide as the main active ingredient circulated on the market.It has the functions of clearing away heat and detoxifying,and reducing inflammation.It has good clinical effects on upper respiratory tract infection,pneumonia,tonsillitis,dysentery and so on.In view of this new drug delivery system,the risk of medication is not predictable.In order to ensure the drug safety after marketing,targeted toxicity studies must be carried out in the non-clinical phase of drug research.It is very important to evaluate its safety,to discuss the symptoms of adverse reactions,the dosage of toxic reactions,the target organs of toxic reactions,the intensity of toxic reactions,and even the symptoms of subsequent toxic reactions that may occur under high doses of drugs.Evaluating the safety of andrographolide nanosuspension for injection comprehensively and objectively provides necessary technical support for the next clinical trial and even the post-marketing risk assessment.On the basis of the notice of CFDA,in strict accordance with the technical specification for new drug registration requirements,under the management of GLP norms,and concerning andrographolide nanosuspension injection,this corpus has tested the acute toxicity of single intravenous injection dose and long-term toxicity of 12 wk intravenous injection dose,carried out the experiment of general pharmacology central nervous system,and the safety evaluation experiment of local toxicity and immunogenicity,and studied its pharmacological toxicology characteristics and safety,providing clinical support for the following clinical experiments of the new drug.The toxicological experiments were conducted as follows:By Up-and-Down Procedure,a LD₅₀0 of Wistar rats caused by andrographolide nanosuspension was 655.7 mg.kg^-1,and the 95%confidence limit was 500-700 mg.kg^-1.This product had obvious stimulation to the injection site,and may even cause muscle rot at the injection site at high concentration,suggesting that long-term toxicity test research and clinical drug use should be closely observed,and dose and concentration should be strictly controlled.Experimental Study on the Toxicity of Repeated Administration,the administration dose was 24 mg.kg^-1(1.6mg.ml^-1concentration),about 2.1times as much as maximum dose of clinical medication,equivalent dose of rats.During the administration,the animals were generally in good condition,with normal activity,diet,drinking water,hematology index,serum biochemical index,urine test and weight gain normal;The administration dose was 60mg.kg^-1(4.0 mg.ml^-1concentration),about 5.4times as much as maximum dose of clinical medication,equivalent dose of rats.During the administration,the animals were generally in good condition,with normal activity,diet,drinking water,hematology index,serum biochemical index,urine test and weight gain,and the injection site of 15%rats showed slight stimulation effect.Latterly,about 10%of rats showed abnormal symptoms such as convulsion,side lying,tremor and rigidity.Organ indicators and pathological examination revealed that the suspected target organ of toxicity was kidney;The administration dose was 150 mg.kg^-1?10.0 mg.ml^-11 concentration?,about 13.4 times as much as maximum dose of clinical medication,equivalent dose of rats.During the administration,the animals were generally in good condition,with normal activity,diet,drinking water,hematology index,serum biochemical index,urine test and weight gain,all rats occurred significantly stimulate and tail swelling at the injection site;Dosing about 50%rats appeared late convulsions,side,tremor,rigidity and other abnormal symptoms;Viscera index and learned suspicious toxic target organs for kidney pathology testing.The toxicity of repeated administration showed that the NOAEL of andrographolide nanosuspension injection was 24 mg.kg^-1.d^-1.When the drug dose was 60mg.kg^-1,the animals showed slight stimulation effect and toxic reaction.When the dosage was 150mg.kg^-1.d^-1,obvious swelling and other irritant symptoms occurred at the injection site.Some rats began to show obvious side effects and renal toxicity.It is stated that the drug toxicity of this product is dose-dependent,and the injection site stimulation and renal toxicity may be induced when given in large doses.It is suggested that the toxicity of repeated drug administration and clinical drug administration should be closely observed and the dose and concentration of drug administration should be strictly controlled.General Pharmacological Studies?Central Nervous System?,general behaviors,autonomic activities:No significant effect was observed on the general behaviors of BALB/c rats,such as gait and hair,after 20,80 and 160 mg.kg^-1.20mg.kg^-1dose of andrographolide nanosuspension L,M and H dosage groups was administered respectively.And the numbers of autonomic activities of rats were detected by YSL-1A autonomous activity recorder after the administration of 20mg.kg^-1.There was no significant differences within each point of 10-240min compared with Veh group?P>0.05?.When 80,160 mg.kg^-11 dose was administrated,the number significantly decreased after 10min compared with Veh group?P<0.01?,and no significant difference was found at each time point of 30-240min compared with Veh group?P>0.05?;Hypnotic effects by subliminal dose of barbital sodium:Compared with the Veh group,there was no significant effect on sleep duration and rate of sleep in BALB/c mice by DAS grade analysis and chi-square test?X²test?when 20?80mg.kg^-1dose of andrographolide nanosuspension L,M was administrated.The 160 mg.kg-1 dose of group H had a significant effect on the duration and rate of sleep in BALB/c mice?P<0.05?;Coordinatedmovement:Comparedwiththe Vehgroup,injectionof andrographolide nanosuspension L,M and H had no significant effect on the coordinated movement of BALB/c mice with doses of 20,80 and 160mg.kg^-1?P>0.05?.Through the research on the central nervous system,it is found that this product has a relatively obvious influence on the central nervous system of rodents mice.When given in large doses,it shows symptoms such as decreased autonomic activity and lethargy,and has no obvious influence on the balance ability of animals.Special safety experiments,In vitro hemolysis:No significant hemolytic and coagulative effect was observed in the treatment with clinical intended concentration of0.3mg.ml^-1;Obvious allergization of was observed on guinea pigs and BN rats when maximum dosage was converted to the concentration of 4mg.ml^-1and 8mg.ml^-1;Vascular stimulation:The intended concentration of 0.3mg.ml^-11 was used clinically.Through this article local toxicity and immunogenicity of observation,found that this product has a mild irritation.Temporary not found obvious hemolysis and allergic symptoms,shows that this new type of drug release systems in improving the class to improve drug allergy rate has a good effect.To sum up,the acute toxicity,blood biochemical parameters,body weight,organ coefficient,central nervous system and irritation to injection site of andrographolide nanosuspension for injection in rats are basically the same as those of the products with andrographolide as the main active ingredient circulated on the market.The toxic target organ is kidney,which is consistent with the literature;In allergic study,the allergic rate of nanosuspension to active allergy in guinea pigs and passive skin allergy in rats was very low,which was in sharp contrast with the high allergic rate reported in literature.It indicated that the allergic rate of nanosuspension to guinea pigs decreased greatly by using new technology.