Skip to main content
Download PDF

Acute toxicological evaluation of AT-533 and AT-533 gel in Sprague-Dawley rats

BMC Pharmacology and Toxicology volume 24, Article number: 54 (2023) Cite this article

Abstract

Background

AT-533 is a novel heat shock protein 90 inhibitor that exerting anti-inflammatory, antiviral, and antitumor efficacy. Furthermore, the gel made of AT-533 as raw material named AT-533 gel has the function of repairing keratitis and dermatitis caused by herpes virus infection. However, the acute safety evaluation of AT-533 and AT-533 gel has not been conducted.

Methods and results

Herein, we performed acute toxicological studies of AT-533 and AT-533 gel in Sprague-Dawley rats. Fifteen-day acute toxicity study of AT-533 was conducted in both male and female Sprague-Dawley rats at doses of 5, 50, 250 and 500 mg/kg and AT-533 gel at 5 g/kg in the study. During experiment, food consumption and mortality were observed and body weight, hematology, serum biochemistry and histopathological assessment of rats were carried out. No abnormal changes were observed in rats percutaneously treated with AT-533 at 5 mg/kg and 50 mg/kg and AT-533 gel. However, loss of appetite and body weight, adverse reactions, toxicologically relevant alterations in hematology and biochemistry were found in rats percutaneously treated with AT-533 at 250 mg/kg and 500 mg/kg during 15-day acute dermic toxicity study.

Conclusions

The aforementioned results suggested that the LD50 of AT-533 is 228.382 mg/kg and the LD50 of AT-533 gel is greater than 5 g/kg. These findings indicated that AT-533 is non-toxic in rats when the dose less than 50 mg/kg and AT-533 gel can be considered a gel with no toxicity at doses less than 5 g/kg.

Peer Review reports

Introduction

Heat shock protein (HSP) was first discovered by Italian scientist Ferruccio Ritossa in the 1960s [1].Based on their molecular weight, HSPs are classified into five families: HSP110 (HSPH), HSP90 (HSPC), HSP70 (HSSPA), HSP60 (HSPD), and small HSPs (e.g., HSPB1 ) [2]. It is a molecular chaperone of 90 kDa that binds to and activates client proteins in ATP-dependent pathways [3] .Chaperones such as HSP90 assist the conformational maturation, folding, and refolding of client proteins during stress [4] .It regulates multiple critical cellular processes through its interactions with them, including cell proliferation, cell cycle, signaling transduction, and tumor progression. Hsp90 expression is increased in several types of cancer, including breast, ovary and prostate cancers, glioblastoma, melanoma, and hepatocellular carcinomas [5].It is known that cancer cells need HSP90 to maintain their homeostasis, where mutated or overexpressed oncoproteins are stabilized. Therefore, HSP90 is an attractive therapeutic target for cancer treatment.

As HSPs rise, some viruses can take advantage of the activated cellular stress response and increase their replication. The E protein of dengue virus (DENV) interacts with HSP90, resulting in a decrease in cellular E protein and an increase in viral titers [6]. HSP90 is a chaperone of various key regulators related to cell growth and survival [7] and used as a therapeutic target for the treatment of cancer [7,8,9], virus [10], autoimmune and inflammatory diseases [11].

At least 18 Hsp90 inhibitors have entered clinical trials targeting NTD since the discovery of the first inhibitor, geldanamycin, in 1994 [12]. There are multiple promising Hsp90 inhibitors in clinical trials, including Onalespib [5].

In latest ten years, we have synthesized a series of 2-aminobenzamide derivatives as novel potent Hsp90 inhibitors, including AT-533 (SNX-25a), SNX-7081 and BJ‑B11 [13,14,15]. All of them are novel analogue of SNX-2112 [16] and has been optimized through structure-activity relationship (SAR) exploration to achieve high Hsp90 affinity [13, 15]. In our previous studies, AT-533 was demonstrated to have many therapeutic potential, including antitumor, antiviral and anti-inflammatory properties [13, 17,18,19]. For example, AT-533 has broad spectrum and high selectivity in inhibiting tumorigenic properties of various cancer cells [20], and inhibits breast cancer growth through HIF-1α/VEGF/VEGFR-2-mediated angiogenesis. Moreover, AT-533 reduced HSV‑1‑induced the nuclear translocation of NF-κB and NLRP3 inflammasome activation via inhibition of the chaperone function ofHSP90 [19]. In addition, AT-533 plays an inhibitory effect on human herpes simplex virus type 1 (HSV‑1) by blocking HSV‑1 nuclear egress and assembly [18] and preventing HSV-1 replication and entry in vitro [21, 22]. Furthermore, AT-533 gel, the gel made of AT-533, has been proved efficiently ameliorating HSV‑1 infected keratitis in a rabbit keratitis model [13] and HSV‑1‑induced skin lesions in C57BL/6 mouse zosteriform [23].

Besides the effectiveness and quality, safety is also an important factor in the development of new drugs [24]. Adverse effects are the major challenge in drug development, and in vivo studies evaluate the toxicity of new therapeutic drugs are critical to support research aimed at evaluating the safety and efficacy of randomized controlled clinical trials in humans. As a compound with varied therapeutic potential, no acute toxicity effect is discussed in AT-533 and AT-533 gel treatment in animal models. In the present study, acute toxicological experiments of AT‑533 and AT‑533 gel were carried out on Sprague-Dawley (SD) rats in accordance with the relevant guiding principles [24], in order to determine the non‑toxic dosage and potential acute toxicity in rats. The present study provides a useful reference for further preclinical research into the use of AT‑533 gel in animals and its clinical application as a skin medication for humans.

Materials and methods

Test substances

AT-533 (Lot No. 20180520) and AT-533 gel (Lot No. 20180706) were acquired according to previous reports [25, 26]. AT‑533 was dissolved in propyleneglycol, PEG400 and sterile water for skin absorption. All solvents meet the requirements of the Chinese Pharmacopoeia [27].

Animals and experimental procedure

All animal experiments were carried out in accordance with the National Institutes of Health’s Guide for the Care and Use of Laboratory Animals. All animal protocols were approved by the Animal Experiment Center of Jinan University. Ninety specific pathogen‑free (SPF) male and female 6-8-week old SD rats were obtained from the Experimental Animal Center of Southern Medical University (Guangzhou, China) under the license number SCXK(YUE)20160041. Feeding conditions are consistent with what we described earlier [26]. Simply, every 5 rats was kept in a plastic cage with labels attached for identification. The rats were housed in an animal room with a 12/12 h light/dark cycle under standard conditions of temperature (21–25℃) and humidity (60–80%) and had free access to food and water. After ten days of acclimation, the fur (about 16 cm2) of the rats from the back to the abdomen were removed by pet hair shaver. Then, the depilatory cream was uniformly applied to the area where the hair had been removed to take off the hair thoroughly. After that, the rats was put into the cage for overnight and was recovered from the stimulation of the depilating cream. The next day, for testing AT-533, 60 rats were randomly divided into 6 groups, including the control, vehicle, AT‑533 (5, 50, 250 and 500 mg/kg). The rats in the control group were administered with sterile saline and the rats in the vehicle group were administered with solvents, respectively. Rats in AT-533 group were transdermally given a corresponding dose of the AT-533. For AT-533 gel, 30 rats were randomly assigned to control group, blank gel group, 5 g/kg AT-533 gel group. Rats in corresponding group were transdermally received sterile saline, blank gel and AT-533 gel, respectively. A total of 5 female rats and 5 male rats were assigned to each group. All rats were transdermally treated with solvent or test substance one time. The test substance was applied directly to the skin and exposure was for 24 h. Then, the test substance was wiped off using warm water.

Observation and detection indicators

All rats were immediately observed after AT-533 or AT-533 gel administration, the animal’s reaction can be observed three times on the day of administration, and then at least once a day for 15 consecutive days of observation period to record any delayed toxic effects. Daily observation records included body weight, skin, fur colour, eyes, autonomic activity, diet, defecation, behaviour of the central nervous system, survival and so on. On day 16 after administration, all rats were anesthetized with pentobarbital sodium (60 mg/kg, i.p.), and blood samples from abdominal aorta were collected for blood routine and biochemical examinations. After that, gross necropsy and anatomy were performed. All the vital organs such as heart, liver, spleen, lung, kidneys and brain were separated and weighed. The organ coefficients (organ-to-final-body-weight ratios) were calculated. Finally, the median lethal dose (LD50) (mg/kg) of AT-533 and AT-533 gel was calculated depended on the number of rats that died during the 15-day acute toxicity test.

Hematological and biochemical parameters

In this study, hematological examination items include white blood cell count (WBC), erythrocyte count (RBC), hemoglobin concentration (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red cell distribution width (RDW), platelet count (PLT) and mean platelet volume (MPV) norepinephrine (NE), lymphocyte (LY), eosinophil (EO), basophilic granulocytes (BA) and monocytes (MO).

Biochemical indexes include total serum protein (TP), albumin (ALB), aminotransferase (ALT), serum aspartate aminotransferase (AST), total bilirubin (T-Bil), direct bilirubin (D-Bil), indirect bilirubin (I-Bil), alkaline phosphatase (ALP), creatinine (CRE), urea nitrogen (BUN), uric acid (UA), triglycerides (TG), blood glucose (GLU), total cholesterol (CHO), high-density lipoprotein cholesterol (HDL), low density lipoprotein-cholesterol (LDL), sodium (Na), potassium (K), and chloride (Cl).

Histopathological assessment

The following vital organs were obtained during necropsy: brain, heart, liver, lung, spleen, kidney and skin. Histological examination for the present study was similar to the procedure in our previous study [28]. Briefly, fresh tissue (1.5 × 1.5 × 0.5 cm) was fixed in 4% paraformaldehyde for 8 h at room temperature, dehydrated using an alcohol gradient, and embedded in paraffin. The tissue was then sectioned into 3-µm slices and stained with hematoxylin and eosin (H&E) and assessed for histopathology using a light microscope (Nikon Corporation, Tokyo, Japan). Besides, calculations were made of the organ coefficients (organ-to-final body-weight) before fixation.

Statistical analysis

All data was analyzed using GraphPad Prism 6.02 (Graph Pad Software, La Jolla, CA, USA) and Microsoft Excel 2013 and presented as mean ± SD. The median lethal dose (LD50) (mg/kg) was calculated by Karber’s method [29], whose basic operation formula is as follows: lgLD50=∑1/2(Xi + Xi + 1)(P i + 1-Pi). Among them, Xi means dose logarithm and Pi means mortality rate. Statistical significance was determined using one-way ANOVA (and nonparametric) followed by Dunnett’s Multiple Comparison Test. The significant difference was indicated by P values less than 0.05, 0.01, 0.001 or 0.0001.

Results

Laboratory observation, food consumption and body weight

In the acute toxicity test of AT-533, 5 deaths (1 male and 4 females) of 250 mg/kg group and 8 deaths (4 males and 4 females) of 500 mg/kg group were recorded during the study. Accordingly, the LD50 of AT-533 calculated using the Karber’s method [29] was 228.382 mg/kg. In addition, rats in control group, vehicle group, 5 mg/kg group and 50 mg/kg group showed a normal diet after transdermal administrating with AT-533. On the contrary, rats in 250 mg/kg group and the 500 mg/kg group showed appetite loss when transdermal treated with AT-533 for 1 to 5 days, and returned to normal on the 6th day (Fig. 1A&B). Correspondingly, the body weight of rats showed the same floating trend (Fig. 1C&D).

Fig. 1
figure 1

Changes of food consumption and body weight of SD rats of different doses of AT-533. (A) Average daily food consumption of female rats. (B) Average daily food consumption of male rats. (C) Average body weight of female rats. (D) Average body weight of male rats. Data presented as mean ± SD (n = 5). A separate one‑way

ANOVA analysis was performed for each time point. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 vs. vehicle group

Full size image

In the acute toxicity test of AT-533 gel, no death was recorded and no abnormal symptom was observed during the study. Thus, LD50 of transdermal administration with single dose of AT-533 gel was greater than 5 g/kg. Normal body weight and food consumption were measured in male and female of three groups (Fig. 2).

Fig. 2
figure 2

Changes in food consumption and body weight of SD rats after transdermal administration of a single dose of AT-533 gel (5 g/kg). (A) Average daily food consumption of female rats. (B) Average daily food consumption of male rats. (C) Average body weight of female rats. (D) Average body weight of male rats. Data presented as mean ± SD (n = 5)

Full size image

Hematology and biochemical examination

When compared with the control group, there was no significant difference in hematology indexes between the vehicle group and the control group. From the results of hematology examinations, the PLT of rats in 500 mg/kg group was remarkably higher than vehicle group rats. In addition, there was no significant difference in hematology index among rats in the vehicle group, 5 mg/kg group, 50 mg/kg group and 250 mg/kg group (Table 1). From the results of biochemical examinations, no significant difference in biochemical items between rats in the vehicle group and the control group was observed. However, ALP for rats of 250 mg/kg group and ALP, CHO, HDL and K+ for rats of 500 mg/kg group were remarkably higher than those of vehicle group rats (Table 2).

Table 1 Hematological values of rats treated with AT-533
Full size table

Values are mean ± SD for 10 rats in each group. aData from 5 rats, b data from 2 rats. *Statistically significant compared to vehicle (p<0.05).

Table 2 Serum biochemical values of rats treated with AT-533
Full size table

In the acute toxicity test of AT-533 gel, the hematology and biochemical results showed no significant differences among control group, blank gel group, AT-533 gel (5 g/kg) group (Tables 3 and 4).

Table 3 Hematological values of rats treated with single dose of AT-533 gel (5 g/kg)
Full size table
Table 4 Serum biochemical values of rats treated with AT-533 single dose of AT-533 gel (5 g/kg)
Full size table

Absolute and relative weight of organs

Furthermore, the absolute organ weight and organ coefficients in female and male rats were shown in Tables 5 and 6, which showed no significant difference between the experimental groups (5 mg/kg and 50 mg/kg) and the vehicle group. Since only one female rat survived in 250 mg/kg group and 500 mg/kg group, this part of data was for reference only and was not discussed anymore.

Table 5 Absolute and relative weights of organs of female rats treated with AT-533
Full size table
Table 6 Absolute and relative weights of organs of male rats treated with AT-533
Full size table

In the acute toxicity test of AT-533 gel, there was no significant difference in organ weight and organ coefficients in female and male rats (Tables 7 and 8). These results suggested that 5 g/kg AT-533 gel was non-toxic to SD rats.

Table 7 Absolute and relative weights of organs of female rats treated with AT-533 gel
Full size table
Table 8 Absolute and relative weights of organs of male rats treated with AT-533 gel
Full size table

Histopathological examination

Histopathological observations were conducted for organs and tissues of all 15-day toxicity portion rats. There were no pathological lesions on heart, liver, spleen, lung, kidney and shin (dosing site) of female and male rats of 5 g/kg AT-533 gel (Fig. 3). Therefore, these results suggested that AT-533 gel used in this test have no damage effect on internal organs.

Fig. 3
figure 3

Histological analysis of heart, liver, spleen, lung, kidneys and skin (dosing site) sections from rats treated with 5 g/kg AT‑533 gel in the 15‑day acute toxicity (H&E staining, x 10). Representative micrographs from (A) female and (B) male rats under the conditions of control, blank gel and AT‑533 gel

Full size image

Discussion

As previously pointed out, AT-533 is first synthesized in our laboratory and in the last few years, there has been an expansion of pharmacological studies carried out with this compound, but there still few have addressed toxicological parameters in animal models. As an important part of the development of new drugs, it’s necessary to preform drug safety evaluation. Thus, this study provided the results of 15-day acute toxicity of AT-533 and AT-533 gel in SD rats.

Body weight changes to some extent indicate adverse reactions of drugs [30,31,32]. In the acute study of AT-533, rats of 5 and 50 mg/kg treated group had normal diet and body weight. However, the food intake, water consumption or weight of rats in the 250 mg/kg and 500 mg/kg treatment group were affected. On the third day of observation, a total of three female rats died at the high doses (250 and 500 mg/kg), indicating that the high doses (250 and 500 mg/kg) AT-533 were toxic and mainly affecting females. The main symptoms of these rats were loss of appetite and diarrhea. No obvious abnormalities were found in other organs except for flatulence, which might be caused by the adverse reaction of AT-533 in stomach. The possible reason maybe the rats licked each other after transdermal administration with AT-533 that cause some drugs reaching stomach through the mouth. From the result, the LD50 of AT-533 calculated is 228.382 mg/kg. Our previous in vitro and in vivo studies showed that the effective concentration was much lower than the LD50 of AT-533 [18, 19, 23], which confirmed the potential druggability of AT-533. Besides, no death, abnormal diet and body weight were found in rats transdermally treated with 5 g/kg AT‑533 gel.

As hematological parameters are key indicators of physiological and pathological status [33], the blood samples were collected to detected to indicate evaluate the toxic effects of the AT-533 and AT-533 gel. Although there was a significant decreased level of lymphocytes (LY) in rats of 50 mg/kg AT-533 treatment group, it was not considered toxic-related since the value of them stayed within the normal reference laboratory range [34]. The increased PLT in rats of 500 mg/kg AT-533 treatment group maybe result from the side effect of AT-533, but the result from only 2 rats, which may be caused by individual differences. Taking into account individual differences, since only 1 ~ 2 rats survived in AT-533 of 250 mg/kg group and 500 mg/kg group, the biochemical parameters and organ coefficients data were for reference only and were not discussed anymore.

In addition, all acquired hematological parameters of rats treated with 5 g/kg AT-533 gel did not show significant variation among rats of control and blank gel groups, which indicated the possible absence of hematological toxicity. The liver and kidneys are target organs to eliminate of xenobiotics, so their function tests are indispensable. The biochemical examinations of rats treated with 5 mg/kg, 50 mg/kg AT-533 or 5 g/kg AT-533 gel reveal no hepatotoxicity and renal toxicity. Organ weight and organ coefficients are important indicators for evaluating the toxic effects of test substances in preclinical toxicology studies of new drugs, and have a high reference value for target organ confirming drug toxicity [24]. The results of Tables 5, 6, 7 and 8 showed no statistical difference among the groups. Furthermore, both gross and histopathological findings showed normal heart, liver, spleen, lung, kidneys and skin morphology of rats treated with 5 g/kg AT-533 gel.

In summary, when SD rats were treated with AT-533 at doses of 5 mg/kg and 50 mg/kg and AT-533 gel at a dose of 5 g/kg, it did not cause mortality. Additionally, the blood routine, biochemical parameters and organ physiological conditions were normal, which indicates that the safe and worthy of AT-533 and its gel to further promote and develop. Therefore, the LD50 of AT-533 is 228.382 mg/kg and the single dose dermic LD50 of AT-533 gel for rats is in excess of 5 g/kg. The aforementioned results provide a reference for subsequent subchronic and chronic toxicity tests of AT‑533 and AT‑533 gel.

Data availability

All the necessary data used to support the results of this study are included in the manuscript.

Abbreviations

ALB:

Albumin

ALP:

Alkaline phosphatase

ALT:

Aminotransferase

AST:

Serum aspartate aminotransferase

BA:

Basophilic granulocytes

BUN:

Urea nitrogen

CHO:

Total cholesterol

Cl:

Chloride

CRE:

Creatinine

D-Bil:

Direct bilirubin

EO:

Eosinophil

GLU:

Blood glucose

HCT:

Hematocrit

HDL:

High-density lipoprotein cholesterol

HGB:

Hemoglobin concentration

Hsp90:

Heat shock protein 90

HSV‑1:

Human herpes simplex virus type 1

I-Bil:

Indirect bilirubin

K:

Potassium

LD50 :

The median lethal dose

LDL:

Low density lipoprotein-cholesterol

LY:

Lymphocyte

MCH:

Mean corpuscular hemoglobin

MCHC:

Mean corpuscular hemoglobin concentration

MCV:

Mean corpuscular volume

MO:

Monocytes

MPV:

Mean platelet volume

Na:

Sodium

NE:

Norepinephrine

PLT:

Platelet count

RBC:

Erythrocyte count

RDW:

Red cell distribution width

SD:

Sprague-Dawley

T-Bil:

Total bilirubin

TG:

Triglycerides

TP:

Biochemical indexes include total serum protein

UA:

Uric acid

WBC:

White blood cell count

References

  1. Ritossa F. Discovery of the heat shock response. Cell Stress Chaperones. 1996;1(2):97–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Lebret T, Watson RW, Molinie V, O’Neill A, Gabriel C, Fitzpatrick JM, Botto H. Heat shock proteins HSP27, HSP60, HSP70, and HSP90: expression in bladder carcinoma. Cancer. 2003;98(5):970–7.

    Article  CAS  PubMed  Google Scholar 

  3. Zhang L, Pitcher LE, Prahalad V, Niedernhofer LJ. Robbins, P. D. recent advances in the discovery of senolytics. Mech Ageing Dev. 2021;200:111587.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Wang X, An D, Wang X, Liu X, Li B. Extracellular Hsp90alpha clinically correlates with Tumor malignancy and promotes migration and invasion in esophageal squamous cell carcinoma. Onco Targets Ther. 2019;12:1119–28.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Cools R, Vermeulen K, Narykina V, Leitao RCF, Bormans G. Radiosynthesis and preclinical evaluation of [(11)C]SNX-ab as an Hsp90alpha,beta isoform-selective PET probe for in vivo brain and tumour imaging. EJNMMI Radiopharm Chem. 2023;8(1):2.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Royle J, Ramirez-Santana C, Akpunarlieva S, Donald CL, Gestuveo RJ, Anaya JM, Merits A, Burchmore R, Kohl A. ;Varjak, M. Glucose-Regulated Protein 78 interacts with Zika Virus Envelope Protein and contributes to a productive Infection. Viruses 2020, 12 (5).

  7. Gomez-Monterrey I, Sala M, Musella S, Campiglia P. Heat shock protein 90 inhibitors as therapeutic agents. Recent Pat Anti-cancer Drug Discov. 2012;7(3):313–36.

    Article  CAS  Google Scholar 

  8. Whitesell L, Lindquist SL. HSP90 and the chaperoning of cancer. Nat Rev Cancer. 2005;5(10):761–72.

    Article  CAS  PubMed  Google Scholar 

  9. Huang KH, Veal JM, Fadden RP, Rice JW, Eaves J, Strachan JP, Barabasz AF, Foley BE, Barta TE, Ma W, Silinski MA, Hu M, Partridge JM, Scott A, DuBois LG, Freed T, Steed PM, Ommen AJ, Smith ED, Hughes PF, Woodward AR, Hanson GJ, McCall WS, Markworth CJ, Hinkley L, Jenks M, Geng L, Lewis M, Otto J, Pronk B, Verleysen K, Hall SE. Discovery of novel 2-aminobenzamide inhibitors of heat shock protein 90 as potent, selective and orally active antitumor agents. J Med Chem. 2009;52(14):4288–305.

    Article  CAS  PubMed  Google Scholar 

  10. Wang Y, Jin F, Wang R, Li F, Wu Y, Kitazato K, Wang Y. HSP90: a promising broad-spectrum antiviral drug target. Arch Virol. 2017;162(11):3269–82.

    Article  CAS  PubMed  Google Scholar 

  11. Tukaj S. Anti-Hsp90 therapy in autoimmune and inflammatory Diseases: a review of preclinical studies. Cell Stress Chaperones. 2016;21(2):213–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Dernovsek J, Zajec Z, Durcik M, Masic LP, Gobec M, Zidar N, Tomasic T. Structure-activity relationships of Benzothiazole-based Hsp90 C-Terminal-domain inhibitors. Pharmaceutics. 2021;13:8.

    Article  Google Scholar 

  13. Xiang YF, Qian CW, Xing GW, Hao J, Xia M, Wang YF. Anti-herpes simplex virus efficacies of 2-aminobenzamide derivatives as novel HSP90 inhibitors. Bioorg Med Chem Lett. 2012;22(14):4703–6.

    Article  CAS  PubMed  Google Scholar 

  14. Liu K, Chen J, Yang F, Zhou Z, Liu Y, Guo Y, Hu H, Gao H, Li H, Zhou W, Qin B, Wang Y. BJ-B11, an Hsp90 inhibitor, constrains the Proliferation and Invasion of Breast Cancer cells. Front Oncol. 2019;9:1447.

    Article  PubMed  PubMed Central  Google Scholar 

  15. Ju H-Q, Xiang Y-F, Xin B-J, Pei Y, Lu J-X, Wang Q-L, Xia M, Qian C-W, Ren Z, Wang S-Y, Wang Y-F, Xing G-W. Synthesis and in vitro anti-HSV-1 activity of a novel Hsp90 inhibitor BJ-B11. Bioorg Med Chem Lett. 2011;21(6):1675–7.

    Article  CAS  PubMed  Google Scholar 

  16. Chandarlapaty S, Sawai A, Ye Q, Scott A, Silinski M, Huang K, Fadden P, Partdrige J, Hall S, Steed P, Norton L, Rosen N, Solit DB. SNX2112, a synthetic heat shock protein 90 inhibitor, has potent antitumor activity against HER kinase-dependent cancers. Clin Cancer Research: Official J Am Association Cancer Res. 2008;14(1):240–8.

    Article  CAS  Google Scholar 

  17. Zhang PC, Liu X, Li MM, Ma YY, Sun HT, Tian XY, Wang Y, Liu M, Fu LS, Wang YF, Chen HY. Liu, Z. AT-533, a novel Hsp90 inhibitor, inhibits Breast cancer growth and HIF-1alpha/VEGF/VEGFR-2-mediated angiogenesis in vitro and in vivo. Biochem Pharmacol. 2020;172:113771.

    Article  CAS  PubMed  Google Scholar 

  18. Li F, Jin F, Wang Y, Zheng D, Liu J, Zhang Z, Wang R, Dong D, Zheng K, Wang Y. Hsp90 inhibitor AT-533 blocks HSV-1 nuclear egress and assembly. J Biochem. 2018;164(6):397–406.

    CAS  PubMed  Google Scholar 

  19. Li F, Song X, Su G, Wang Y, Wang Z, Qing S, Jia J, Wang Y, Huang L, Zheng K, Wang Y. AT-533, a Hsp90 inhibitor, attenuates HSV-1-induced inflammation. Biochem Pharmacol. 2019;166:82–92.

    Article  CAS  PubMed  Google Scholar 

  20. Wang S, Wang X, Du Z, Liu Y, Huang D, Zheng K, Liu K, Zhang Y, Zhong X, Wang. Y. SNX-25a, a novel Hsp90 inhibitor, inhibited human cancer growth more potently than 17-AAG. Biochem Biophys Res Commun. 2014;450(1):73–80.

    Article  CAS  PubMed  Google Scholar 

  21. Qin S, Hu X, Lin S, Xiao J, Wang Z, Jia J, Song X, Liu K, Ren Z, Wang Y. Hsp90 inhibitors prevent HSV-1 replication by directly targeting UL42-Hsp90 complex. Front Microbiol. 2021;12:797279.

    Article  PubMed  Google Scholar 

  22. Song X, Wang Y, Li F, Cao W, Zeng Q, Qin S, Wang Z, Jia J, Xiao J, Hu X, Liu K, Wang Y, Ren Z. Hsp90 inhibitors inhibit the entry of herpes Simplex Virus 1 into Neuron cells by regulating cofilin-mediated F-Actin reorganization. Front Microbiol. 2021;12:799890.

    Article  PubMed  Google Scholar 

  23. Wang Y, Wang R, Li F, Wang Y, Zhang Z, Wang Q, Ren Z, Jin F, Kitazato K, Wang Y. Heat-shock protein 90alpha is involved in maintaining the stability of VP16 and VP16-mediated transactivation of alpha genes from herpes simplex virus-1. Mol Med. 2018;24(1):65.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. SFDA Technical guidelines. of acute toxicity testing for chemical drugs. SFDA Guidelines No. [H] GPT1-1 2005.

  25. Wu Y, Li M, Guo Y, Liu T, Zhong L, Huang C, Ye C, Liu Q, Ren Z, Wang Y. The effects of AT-533 and AT-533 gel on liver cytochrome P450 enzymes in rats. Eur J Drug Metab Pharmacokinet. 2022;47(3):345–52.

    Article  CAS  PubMed  Google Scholar 

  26. Wu Y, Song X, Qin S, Chen P, Huang L, Wang Q, Shan T, Liang F, Liao X, Liu Q, Huang Y, Wang Y. Subacute toxicological evaluation of AT-533 and AT-533 gel in Sprague-Dawley rats. Exp Ther Med. 2021;21(6):632.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Commission, C. P. Chinese pharmacopoeia. Beijing: China Medical Science and Technology Press; 2020.

    Google Scholar 

  28. Wu Y, Jin F, Wang Y, Li F, Wang L, Wang Q, Ren Z, Wang Y. In vitro and in vivo anti-inflammatory effects of theaflavin-3,3’-digallate on lipopolysaccharide-induced inflammation. Eur J Pharmacol. 2017;794(1):52–60.

    Article  CAS  PubMed  Google Scholar 

  29. Tan P. Application of Excel software to calculate the median lethal dose. J Shangxi Med Univ. 2010;41(10):914–6.

    Google Scholar 

  30. Deyno S, Abebe A, Tola MA, Hymete A, Bazira J, Makonnen E, Alele PE. Acute and sub-acute toxicity of Echinops kebericho decoction in rats. BMC Complement Med Ther. 2020;20(1):1–11.

    Article  Google Scholar 

  31. Dong Z, Tang S-s, Li C-h, Tang Z-s, Yang Z. -h.;Zeng, J.-g. Safety assessment of MPTA: an oral acute and 90-day sub-chronic toxicity study in Sprague-Dawley rats. Regul Toxicol Pharmacol 2022, 105188.

  32. Li F, Wang L, Cai Y, Luo Y, Shi X. Safety assessment of desaminotyrosine: Acute, subchronic oral toxicity, and its effects on intestinal microbiota in rats. Toxicol Appl Pharm. 2021;417:115464.

    Article  CAS  Google Scholar 

  33. Deyno S, Abebe A, Tola MA, Hymete A, Bazira J, Makonnen E, Alele PE. Acute and sub-acute toxicity of Echinops kebericho decoction in rats. BMC Complement Med Ther. 2020;20(1):2.

    Article  PubMed  PubMed Central  Google Scholar 

  34. yali W. r. W. t. W. h. T. j. g. b. Z. Establishment of routine blood reference intervals in SD rats. Chin J Med Device. 2015;28(07):11–5.

    Google Scholar 

Download references

Acknowledgements

We would like to thank Dr. Pengxiao Chen (GuangZhou (Jinan) Biomedical Research and Development Center Co. Ltd) for technical guidance during the animal experiments and Mr. Hui Chen (Guangzhou University of Chinese Medicine) for the care and encouragement of the experimental progress.

Funding

This work was supported by the Science and Technology Foundation of Shenzhen (JCYJ20200109144410181) and the National Natural Science Foundation of China (82273040), Natural Science Foundation of Guangdong Province (2019A1515010046) and Guangzhou Major Program of the Industry‑University‑Research collaborative innovation (201704030087).

Author information

Author notes

  1. Lishan Zhong and Yanting Wu contributed equally to this work.

Authors and Affiliations

  1. Department of Cell Biology, College of Life Science and Technology, Jinan University, Guangzhou, China

    Lishan Zhong, Yanting Wu, Chen Huang, Cui-fang Ye, Zhe Ren & Yifei Wang

  2. Guangdong Province Key Laboratory of Bioengineering Medicine, Guangzhou, China

    Lishan Zhong, Yanting Wu, Chen Huang, Cui-fang Ye, Zhe Ren & Yifei Wang

  3. Guangdong Provincial biotechnology drug and Engineering Technology Research Center, Guangzhou, China

    Lishan Zhong, Yanting Wu, Chen Huang, Cui-fang Ye, Zhe Ren & Yifei Wang

  4. Guangzhou (Jinan) Biomedical Research and Development Center Co. Ltd, Guangzhou, China

    Yanting Wu & Yifei Wang

  5. Guangdong Provincial Clinical Research Center for Geriatrics, Shenzhen Clinical Research Center for Geriatrics,Shenzhen People’s Hospital, The Second Clinical Medical College, Jinan University, Shenzhen, Guangdong, China

    Kaisheng Liu

  6. Shenzhen People’s Hospital, The First Affiliated Hospital, Southern University of Science and Technology, Shenzhen, Guangdong, China

    Kaisheng Liu

Authors
  1. Lishan Zhong
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yanting Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Chen Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kaisheng Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Cui-fang Ye
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Zhe Ren
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yifei Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Yanting Wu and Yifei Wang designed the study; Yanting Wu drafted the manuscript; Lishan Zhong, Yanting Wu, Chen Huang, Kaisheng Liu, Cuifang Ye, Zhe Ren performed the experiments; Lishan Zhong analyzed the data; Yanting Wu and Yifei Wang reviewed the final version of the paper; Kaisheng Liu and Yifei Wang provide funding support. All the authors read and approved the final version of the manuscript.

Corresponding authors

Correspondence to Kaisheng Liu or Yifei Wang.

Ethics declarations

Ethics approval and consent to participate

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed and the study protocols were approved by The Institutional Animal Care and Use Committee of Jinan University Guangzhou, PR China (Approval Number: IACUC-2020-000038). The rats were not subjected to any needless, painful, or frightening manipulations. Before being sacrificed, all rats were anesthetized with pentobarbital sodium to avoid pain and suffering and the blood samples and organs of sacrificed rats were gained following the laboratory standards of Jinan University Guangzhou, PR China. The study was reported in accordance with ARRIVE guidelines.

Consent for publication

Not applicable.

Competing interests

The authors declare no conflicts of interest.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhong, L., Wu, Y., Huang, C. et al. Acute toxicological evaluation of AT-533 and AT-533 gel in Sprague-Dawley rats. BMC Pharmacol Toxicol 24, 54 (2023). https://doi.org/10.1186/s40360-023-00696-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s40360-023-00696-5

Keywords

BMC Pharmacology and Toxicology

ISSN: 2050-6511

Contact us