Etanercept inhibits the in vitro activity of human TNF. It modulates biological responses induced or regulated by TNF, such as the expression of adhesion molecules (E-selectin, ICAM-1) and the production of interleukin-6 (IL-6), matrix metalloproteinase-3 (MMP-3), and IL-1.[3]

The mean arthritis scores and radiographic scores were considerably lowered by etanercept (10 mg/kg; subcutaneous injection; every 3 days for 3 weeks) [4].

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Etanercept is efficacious in many in vivo models of inflammation, including arthritis.
In clinical trials for Rheumatoid Arthritis (RA), significant improvements were observed: In patients with an inadequate response to Methotrexate (MTX), adding ETN (25mg twice weekly) led to 71% achieving ACR20 vs 27% with placebo+MTX at 24 weeks, and 39% achieving ACR50 vs 3%.
In early RA, monotherapy with ETN (25mg twice weekly) resulted in 72% of patients having no radiographic erosion progression at 12 months vs 60% with MTX.
In the TEMPO trial for established RA, combination therapy (ETN+MTX) was superior to either agent alone: ACR20 was 85% (combo) vs 76% (ETN) vs 75% (MTX); ACR50 was 69% vs 48% vs 43%; and 48% in the combo group achieved clinical remission.
A once-weekly 50mg dose regimen showed comparable efficacy to the 25mg twice-weekly regimen.
In Psoriatic Arthritis (PsA), ETN (25mg twice weekly) led to 87% of patients meeting PsARC vs 23% on placebo, and 73% achieving ACR20 vs 13% at 12 weeks.[3]

Animal/Disease Models: Sixweeks old male Lewis rat (adjuvant-induced arthritis (AIA) model) [4]
Doses: 10 mg/kg
Route of Administration: Sc; every 3 days for 3 weeks
Experimental Results: Average arthritis score Significant decrease; radiographic score diminished Dramatically at the end of the treatment period.

Metabolism / Metabolites
In biological tissues, V3+ and V4+ predominate due to the reducing environment; however, V5+ is formed in hyperoxic plasma. In biological tissues, V3+ and V4+ predominate due to the reducing environment; however, V5+ is formed in hyperoxic plasma. In calves, after daily oral administration of 10-20 mg/kg ammonium metavanadate, the vanadium content in the liver ranged from 0.3 to 5.1 ppm (wet tissue), and in the kidneys from 6.0 to 40 ppm. In healthy volunteers, after a single subcutaneous injection of 25 mg, the average time to peak serum concentration was 51 hours, and the maximum concentration (Cmax) was 1.46 mcg/mL (range 0.37-3.47). The elimination half-life was approximately 68 hours. A twice-weekly dosing regimen maintained steady-state concentrations. A once-weekly subcutaneous injection of 50 mg showed similar pharmacokinetic characteristics at steady state.
No dose adjustment is required for renal or hepatic impairment, and pharmacokinetic characteristics are similar in men and women, as well as in different age groups (65–87 years vs <65 years).
Concomitant administration of methotrexate (MTX) does not alter the pharmacokinetic characteristics of etanercept. [3]

Toxicity Summary
Identification and Uses: Ammonium vanadate is an orange powder. It is used as a spray chromogenic agent in pharmaceutical analytical toxicology. Human Exposure and Toxicity: No relevant data are currently available. Animal Studies: Addition of ammonium vanadate (10 or 20 mg/L) to drinking water had no effect on the development of colorectal tumors in mice treated subcutaneously with 1,2-dimethylhydrazine (DMH) for 20 weeks. Although the thymidine incorporation was increased, ammonium vanadate had no effect on the incidence or type of DMH-induced tumors. Two months after feeding rats with 15 mg/kg vanadium in the form of ammonium vanadate, increased ventricular pressure and pulmonary hypertension were observed, but no changes were seen in systemic circulation. When rats and mice were orally administered ammonium vanadate at doses of 0.005–1 mg vanadium/kg for 21 days (higher dose) to 6 months (lower dose), a dose of 0.05 mg vanadium/kg was found to be the threshold for inducing dysfunction of conditioned reflexes in rats and mice. Ecotoxicity studies: Ammonium metavanadate at 0.02 mg/L interferes with cell division in the freshwater algae Chlorella pyrenoidosa, while a concentration of 0.25 mg/L is lethal. Identification and uses: Ammonium metavanadate is a white or slightly yellow crystalline powder. It is used for dyeing and printing wool fabrics; wood dyeing; the production of vanadium black and "non-fading inks"; the production of vanadium luster in ceramics; as a developing agent; for microscopic hematoxylin staining; and as an analytical chemical reagent. Because it readily converts to vanadium pentoxide at high temperatures, it is often used as a substitute for vanadium pentoxide. Human exposure and toxicity: A worker was exposed to a large amount of dry ammonium metavanadate dust over a 6-hour period while shoveling the powder into a hopper. Within 2 hours of starting work, the worker reported symptoms including headache behind the eyes, tearing, dry mouth, and a green tongue. A noticeable green discoloration appeared on the skin of the fingers, scrotum, and thighs. The worker was also reported to have nasal congestion and lethargy. The next day, his testicles became swollen and tender. On the third day after exposure, he developed wheezing, dyspnea, and a cough producing green sputum. Over the next two weeks, he experienced multiple episodes of hemoptysis. Wheezing and dyspnea persisted for approximately one month; chest symptoms reached their peak three weeks after the incident. At an examination six weeks after the last exposure, he had no symptoms other than partial obstruction of the left nostril and redness of the nasal mucosa. Chest examination was unremarkable. Pulmonary function assessment showed normal lung volume, forced expiratory flow, and gas exchange. A mild eosinophilic saturation was observed in peripheral blood. In human fibroblast culture, VO3- induces DNA synthesis and cell growth. Ammonium metavanadate was not found to increase the frequency of structural chromosomal aberrations in human leukocytes, but it was found to significantly increase the number of numerical aberrations, micronuclei, and satellite-related chromosomes. Fluorescence in situ hybridization (FISH) of human lymphocytes using α-satellite centromere-specific DNA probes confirmed the aneuploidy pathogenicity of vanadium. Animal experiments: Mice exposed to a 20 mg/kg ammonium metavanadate solution of vanadium developed acute tubular necrosis, pulmonary hemorrhage, and lymphoid tissue necrosis. In a 3-month study, rats were given ammonium metavanadate at a concentration of 200 mg/L (calculated as vanadium) in their drinking water. The animals exhibited slow weight gain and anemia. Gross pathological examination revealed parenchymal malnutrition in the liver and kidneys of some animals, with tubular structures forming within the renal tubules. Acute exposure to vanadium oxides and vanadium salts (including ammonium metavanadate) (oral and subcutaneous injection) in dogs and rabbits has been reported to produce neurophysiological effects, including central nervous system disorders (impaired conditioned reflexes and neuromuscular excitability). Furthermore, the teratogenicity of ammonium metavanadate in hamsters was investigated. Twenty pregnant hamsters in each group were intraperitoneally injected with 0, 0.47, 1.88, and 3.75 mg/kg ammonium metavanadate on days 5 to 10 of gestation. Pregnant female hamsters were sacrificed on day 15. The results showed that the incidence of skeletal deformities in mice was significantly increased and the sex ratio was significantly decreased in all dosage groups. Ammonium metavanadate improved the conversion and recovery rates of Saccharomyces cerevisiae D7 strain; its activity was highest without metabolic activation. Micronucleus assays in the bone marrow of mice after intragastric administration showed a positive result for ammonium metavanadate. Conversely, no difference was found between the control and administration groups in chromosomal structural aberration assays performed at 24 and 36 hours post-administration. In studies using ammonium metavanadate at concentrations of 5-40 uM, the hprt gene in Chinese hamster V79 cells and the gpt site in the hprt-/gpt+ transgenic cell line G12 both showed weak mutagenicity. Female mice were intraperitoneally injected with 2.5, 5, or 10 mg/kg ammonium metavanadate every 3 days for 3, 6, or 9 weeks. Results showed a dose-dependent increase in resistance to lethal E. coli endotoxins over a period of up to 6 weeks, while resistance to lethal Listeria monocytogenes decreased in a dose-dependent manner. Hepatomegaly and splenomegaly, increased splenic megakaryocytes, and increased erythrocyte precursor production were also observed.

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Effects during pregnancy and lactation
◉ Overview of use during lactation
Etanercept is minimally excreted into breast milk and poorly absorbed by infants, consistent with its high molecular weight of approximately 150,000 Da. Waiting at least 2 weeks postpartum before resuming treatment minimizes drug transfer to the infant. There are currently no long-term follow-up data on breastfeeding infants while mothers are taking etanercept. The risk of adverse reactions in older infants is unknown, but is considered unlikely. Most experts and professional guidelines consider the risk to breastfeeding infants to be low and that it can be used during breastfeeding.
◉ Effects on Breastfed Infants
A woman with rheumatoid arthritis started receiving 25 mg of etanercept subcutaneously twice weekly 3 months postpartum, then switched to 50 mg subcutaneously once weekly. Her infant was breastfed until 6 months of age (feeding extent unspecified). The infant was reported to be in good health at age 3.
A case-control study of women with chronic arthritis found that 5 women received etanercept treatment during pregnancy and lactation (feeding extent unspecified). No differences were observed in growth indicators, developmental milestones, vaccinations, or diseases in the 5 infants during their first year of life compared to breastfed infants who were not exposed to the drug.
Six infants were breastfed by mothers with rheumatoid arthritis or ankylosing spondylitis (2 exclusively breastfed, 4 partially breastfed), and these mothers received etanercept treatment during pregnancy and prior to enrollment in the study. One mother reported that her infant experienced a mild rash and high-pitched crying, both of which resolved spontaneously without intervention. All infants had normal growth indicators at a 6-month health checkup.
A nationwide prospective registry study was conducted in Spain on patients with rheumatic diseases receiving DMARDs. Three infants whose mothers were taking etanercept were breastfed (the extent of breastfeeding was not specified), and no mild or severe adverse events were reported in the infants.
◉ Effects on lactation and breast milk
As of the revision date, no relevant published information was found.

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Toxicity Data
LC50 (rat) = 7.8 mg/m3/4h
LC50 (rat) = 340 mg/m3/4h

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Interactions
Vanadium compounds are now commonly added to nutritional supplements and are being developed for the treatment of diabetes. Previous studies have shown that bis(maltol)vanadium oxide (IV) (BMOV) increases tissue uptake compared to vanadium sulfate (VS). Our primary objective was to verify the hypothesis that complexation increases vanadium uptake, and that this effect is independent of oxidation state. A secondary objective was to compare the effects of vanadium complexation and oxidation state on iron, copper, and zinc in tissues. Wistar rats were fed ammonium metavanadate (AMV), vanadium sulfate (VS), or dimethyl vanadate (BMOV) (all at 1.2 mM in drinking water). After 12 weeks, tissue vanadium uptake in the BMOV or AMV groups was significantly higher than in the VS group (p < 0.05). BMOV resulted in decreased tissue zinc content and increased bone iron content. The three compounds were compared in a cellular uptake model (Caco-2 cells). At 10 minutes, vanadium uptake in the VS group was higher than in the BMOV or AMV groups, but vanadium uptake in the BMOV group (only 250 μM, 60 minutes) was significantly higher than in the AMV or VS groups. These results indicate that neither complexation nor oxidation state alone is sufficient to predict relative uptake, tissue accumulation, or trace element interactions. Previous studies in our laboratory have confirmed the potential anticancer effects of vanadium (a dietary micronutrient) in in vivo rat models of liver, colon, and breast cancer. In this paper, we further explored the anticancer effects of this essential trace element by investigating several biomarkers of chemical carcinogenesis, particularly cell proliferation and oxidative DNA damage. We induced liver cancer in male Sprague-Dawley rats by adding 0.05% 2-acetaminofluorene (2-AAF) daily to their basal diet for 5 days a week. Simultaneously, we freely supplemented the rats with vanadium in the form of ammonium metavanadate (0.5 ppm, equivalent to 4.27 μmol/L). Compared with the carcinogen control group, continuous administration of vanadium reduced relative liver weight, nodule incidence (79.99%), total nodule count, and nodule number (p<0.001; 68.17%), and improved hepatocyte structure. Vanadium treatment further restored the activity of hepatic uridine diphosphate (UDP)-glucuronide transferase and UDP-glucose dehydrogenase, inhibited lipid peroxidation, and prevented the formation of glycogen-accumulating precancerous lesions in the initiation-promote model (p<0.01; 63.29%). Long-term vanadium treatment also reduced the bromodeoxyuridine (BrdU) labeling index (p<0.02) and inhibited cell proliferation during hepatocellular precancerous lesions. Finally, short-term vanadium exposure significantly reduced the production of 8-hydroxy-2'-deoxyguanosine (p<0.001; 56.27%), DNA mass aspect ratio (p<0.01), and mean frequency of tailed DNA in the liver of precancerous rats (p<0.001). This study suggests that vanadium may play a role in inhibiting cell proliferation and preventing early DNA damage in vivo. Vanadium has a chemopreventive effect on the early stage of 2-AAF-induced rat hepatocellular carcinoma. Metals, including vanadium (V), have a strong affinity for the sulfhydryl (-SH) groups in biomolecules, including glutathione (GSH) in tissues. Therefore, we further investigated the interaction between ammonium vanadate [NH₄VO₃] and glutathione (a toxic biomarker), and the role of glutathione in the detoxification and binding processes of whole blood components (including plasma and cytoplasmic fractions). We examined the effects of different concentrations of ammonium vanadate [NH₄VO₃] on the levels of reduced glutathione in whole blood components (plasma and cytoplasmic fractions). The results showed that the consumption of glutathione in both plasma and cytoplasmic fractions was positively correlated with the concentration of ammonium vanadate, and the decrease in glutathione levels was more significant with prolonged incubation time. These findings suggest that the changes in glutathione (GSH) levels induced by ammonium metavanadate may be due to the formation of adducts (V-SG) between vanadium and glutathione or increased production of oxidized glutathione (2GSH + V(+5) ⇌ GSSG). This change in GSH metabolic status provides some information about the toxic mechanism of ammonium metavanadate and the protective effect of glutathione. This study examined the expression patterns of heat shock protein (Hsp) 72/73 and glucose-regulated protein (Grp) 94 in the liver, kidney, and testes of rats injected with a sublethal dose of ammonium metavanadate (5 mg/kg/day). Furthermore, to assess the protective effect of green tea, some animals were given a decoction rich in antioxidant compounds as their sole beverage. In control animals, stress protein expression was organ-dependent: anti-Grp94 antibody detected two bands (96 kDa and 98 kDa) in the kidney and liver, but only a 98 kDa band in the testes; anti-Hsp72/73 antibody showed that constitutive Hsp73 was present in all organs, while inducible Hsp72 was present only in the kidney and testes. In the kidneys of vanadium-treated rats, Hsp73 expression was upregulated by approximately 50%, while Hsp72 expression was downregulated by 50-80%. No similar effects were observed in the liver and testes. In the liver and kidneys of vanadium-treated rats, Grp94 expression was upregulated by 50% and 150%, respectively, but no changes were observed in the testes. In rats whose only beverage was green tea, the expression level of the 96 kDa protein in the liver was reduced in both the control and vanadium-treated groups. However, drinking green tea did not prevent vanadium-induced downregulation of Hsp72 expression in the kidneys of vanadium-treated rats. For more complete data on interactions of ammonium metavanadate (10 types), please visit the HSDB record page.

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Non-human toxicity values
Rat inhalation LC50: 0.34 mg/L/4 hr
Rat oral LD50: 162 mg/kg body weight
Rat oral LD50: 58.1 mg/kg
Rat dermal LD50: 2102 mg/kg
Rat intraperitoneal injection LD50: 18 mg/kg
Rat subcutaneous injection LD50: 23 mg/kg
Rat inhalation LC50: 7.8 mg/m³/4 hr>>Infection: In placebo-controlled trials, no increase in the frequency or nature of infection (including serious infection) was observed compared to placebo. However, some observational studies have reported an increased incidence of non-serious respiratory infections.
Tuberculosis (TB): Cases, including miliary tuberculosis, have been reported through post-marketing surveillance. Screening for latent tuberculosis is recommended.
Autoimmune diseases: New antinuclear antibodies (ANA: 11% vs. 5% in placebo) and anti-double-stranded DNA antibodies (anti-dsDNA: 15% vs. 4% in placebo) have been observed. There have been case reports of drug-induced systemic lupus erythematosus (SLE)-like syndromes, with symptom relief upon discontinuation of the drug.
Malignancies/Lymphomas: Registry data show an increased standardized incidence ratio (SIR) for lymphomas in rheumatoid arthritis (RA) patients receiving anti-TNF therapy (SIR 3.8 with or without etanercept), but this may reflect selection bias (more severe disease). Causality has not been established.
Demyomyelinating syndromes: A temporal association has been reported between the occurrence of central nervous system demyelinating diseases (e.g., multiple sclerosis, optic neuritis) and etanercept (ETN) use.
Congestive heart failure (CHF): In prospective trials of patients with New York Heart Association (NYHA) class II-IV CHF, ETN showed no benefit in reducing mortality or CHF hospitalization rates compared to placebo, and in one trial, the ETN group showed a trend toward increased infection rates. [3]

[1]. Etanercept: An overview. J Am Acad Dermatol. 2003;49(2 Suppl):S105-S111.

[2]. Goldenberg MM. Etanercept, a novel drug for the treatment of patients with severe, active rheumatoid arthritis. Clin Ther. 1999;21(1):75-2.

[3]. Etanercept in the treatment of rheumatoid arthritis. Ther Clin Risk Manag. 2007;3(1):99-105.

[4]. Etanercept improves endothelial function via pleiotropic effects in rat adjuvant-induced arthritis. Rheumatology (Oxford). 2016;55(7):1308-1317.

Ammonium metavanadate is a white crystalline powder. Slightly soluble in water, with a density greater than water. Decomposes at 410 °F (210 °C). May release toxic gases. Moderately toxic. Irritating. Used as a drying agent for paints, inks, and dyes. Loses ammonia upon heating.
See also: Etanercept (note moved to).
Etanercept is a dimer fusion protein composed of two extracellular domains of the human 75 kDa TNF receptor (p75) linked to the Fc segment of human IgG1. It is produced using recombinant DNA technology in a mammalian cell expression system.
It is approved for the treatment of rheumatoid arthritis (RA) and psoriatic arthritis (PsA).
It can be used as monotherapy or in combination with methotrexate (MTX), with combination therapy showing superior efficacy in delaying radiographic progression of RA. It can improve the health-related quality of life (HRQOL) and functional status (HAQ) of RA patients, and may reduce the use of medical resources and increase employment. [3]

H4NO3V

116.978161811829

185243-69-0

516859

Colorless to light yellow liquid

200 °C (with decomposition)

1

3

0

5

36.5

0

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)