Absorption, Distribution and Excretion
Approximately 80% of p-toluenesulfonamide is excreted in the urine of rats after administration, with half of the compound being oxidized to p-sulfonylbenzoic acid. Over 90% of p-sulfonylbenzoic acid metabolites are excreted unchanged, but the urine-to-fecal ratio varies considerably among different animals. After oral administration of the 14C-labeled compound to rats, the label is rapidly excreted, primarily in the urine (66-89% of the dose), with only a small amount in the feces (2-8% of the dose). The (14)C in the feces is 4-sulfonylbenzoic acid, which may originate from tissues. Metabolism/Metabolites The purpose of this study was to investigate the in vivo and in vitro metabolism of p-toluenesulfonamide (PTS) and its effects on cytochrome P450 enzymes (CYP450). Total CYP450 and microsomal protein levels were measured in rats after intravenous PTS pretreatment. CYP-specific substrates were incubated with rat liver microsomes. The activities of specific CYP isoenzymes were determined by high-performance liquid chromatography (HPLC). CYP chemical inhibitors were added to the incubation mixture to investigate the major CYP isoenzymes involved in PTS metabolism. The effect of PTS on CYP isoenzymes was investigated by incubating PTS with specific substrates. Groups administered PTS at doses of 33 and 99 mg/kg/day had total CYP contents of 0.66 ± 0.17 and 0.60 ± 0.12 nmol/mg, respectively. K_m and V_max were 92.2 μmol/L and 0.0137 nmol/min/mg protein, respectively. CYP2C7, CYP2D1, and CYP3A2 may be involved in PTS metabolism in rat liver. Sulfadiazine and ketoconazole exhibit mixed mechanisms of inhibition of PTS metabolism, while quinidine inhibits PTS metabolism in a non-competitive manner. PTS has minimal effect on the activity of selected CYP isoenzymes. Overall, PTS is relatively safe when used in combination with other drugs. However, caution should be exercised when PTS is used in combination with CYP inhibitors and substrates of CYP2C, CYP2D, and CYP3A. This study aimed to investigate the metabolism of p-toluenesulfonamide (PTS) in the liver and its interactions with CYP isoenzymes, P-glycoproteins (P-gp), and other drugs. Known CYP substrates, inducers, and inhibitors, as well as P-gp inhibitors, were used, and metabolites were determined using high-performance liquid chromatography (HPLC). Male Wistar rats were pre-administered intraperitoneally with phenobarbital (PB), ketoconazole (Ket), or verapamil (Ver) for 3 days, followed by in situ hepatic perfusion of PTS into the circulatory system. Before liver perfusion with dextromethorphan (Dex) and phenacetin (Phe), rats were pre-administered with PTS (33 mg/kg/day or 99 mg/kg/day) intraperitoneally for 4 days. The effects of PTS on five CYP isoenzymes and the probability of in vitro drug interactions between PTS and chlorotoxin and 5-fluorouracil (5-FU) were investigated using microsomal incubation. At 60 minutes after perfusion, the PTS area in the PB and Ket groups was 61.4% and 133.6% of that in the control group, respectively. PB treatment enhanced PTS metabolism, while Ket treatment inhibited it, indicating that hepatic CYPs are involved in PTS metabolism. PTS pretreatment (dose of mg/kg/day) slowed the metabolism of Dex and Phe, while in vitro incubation experiments did not show any effect of PTS (0-160 μmol/L) on CYP activity. When co-incubated with chlorotoxin, the production of PTS metabolites was 50.7% of that in the negative control group. Further clinical studies are needed to assess the safety of this combination therapy, given the potent inhibitory effect of chlorotoxin on PTS. Fifteen adult rainbow trout were exposed to an aqueous solution of (14)C-toluenesulfonyl chloride sodium (purity 93.7%, specific activity 1.2 μgCi/μM) at a concentration of 20 mg/L (twice the intended treatment concentration) for 1 hour, and then transferred to fresh water. The well water temperature was 11–13 °C. Toluenesulfonyl chloride sodium was rapidly reduced to the major metabolite p-toluenesulfonamide, but its content was not quantified. …50% of the orally administered o-toluenesulfonamide and p-toluenesulfonamide were excreted in the urine and metabolized to o-sulfonylbenzoic acid and p-sulfonylbenzoic acid, respectively. For more complete data on the metabolism/metabolites of p-toluenesulfonamides (6 in total), please visit the HSDB record page.

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Biological Half-Life
Juvenile trout were exposed to 20 mg/L (twice the therapeutic concentration) of cyclic UL-14C-toluenesulfonyl chloride sodium (93.7% purity, specific activity 1.2 μCi/μM) for up to 1 hour, followed by recovery in freshwater to assess the accumulation and distribution of residues in tissues. Well water temperatures ranged from 11.6 to 12.2 °C. Based on radiodating data, the half-life of the p-toluenesulfonamide equivalent in fry was estimated to be 27.3 hours, while the half-life of p-toluenesulfonamide residues in whole fish homogenates was determined by high-performance liquid chromatography (HPLC) to be 36.3 hours. Based on radiodating data, the half-life of p-toluenesulfonamide residues in juvenile fish was estimated to be 32.6 hours, while the half-life of p-toluenesulfonamide residues in whole fish samples was determined by HPLC to be 40.3 hours.

Toxicity Summary
Identification and Uses: p-Toluenesulfonamide is a solid. It is used in organic synthesis, plasticizers and resins, as well as as a bactericide and fungicide in paints and coatings. It has also been used as an experimental therapy. Human Studies: It is known to be a common contact allergen. Animal Studies: In a 90-day feeding study, dogs were fed a mixture of p-toluenesulfonamide (68%) and o-toluenesulfonamide (32%) at doses up to 3000 mg/kg of diet (equivalent to 75 mg/kg body weight/day), and no treatment-related adverse reactions were observed. In a 90-day feeding study, rats were fed diets containing a mixture of p-toluenesulfonamide (68%) and o-toluenesulfonamide (32%) at doses of 0, 300, 1000, or 3000 mg/kg of diet, equivalent to approximately 15, 50, or 150 mg/kg body weight/day, respectively. Results showed that only weight gain and a slight decrease in food consumption were observed in the 3000 mg/kg diet group, which were the only treatment-related effects. Pregnant rats were administered a mixture of p-toluenesulfonamide (68%) and o-toluenesulfonamide (32%) via gavage at doses of 0, 50, 250, or 500 mg/kg body weight, respectively. Results showed that the 250 and 500 mg/kg body weight regimens significantly reduced maternal weight gain during treatment. At the same dose levels, post-implantation embryo loss rate increased in a dose-dependent manner, and fetal weight decreased. No teratogenic effects were observed. The mutagenicity of p-toluenesulfonamide was investigated using the Salmonella Typhimurium/microsomal assay, the Drosophila basal assay, and the mouse micronucleus assay. No mutagenic activity was found in any of these assays.

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Non-human toxicity values
Oral LD50 in rats: 2400 mg/kg body weight / mixture of o-toluenesulfonamide (41%) and p-toluenesulfonamide (51%) /
Oral LD50 in rats: 2330 mg/kg body weight
Oral LD50 in rats: >2000 mg/kg
Oral LD50 in mice: 400 mg/kg
Intraperitoneal LD50 in mice: 250 mg/kg

Toluene-4-sulfonamide is a sulfonamide, which is benzenesulfonamide with a methyl group attached at the 4-position. p-Toluenesulfonamide is a low molecular weight organic compound with potential antitumor activity. After intratumoral injection, p-Toluenesulfonamide increases lysosomal membrane permeability (LMP) and promotes the release of cathepsin B. The lysosomal-released cathepsin B cleaves and activates pro-apoptotic B-cell lymphoma 2 (Bcl-2) family members BH3-interacting domain death agonist (Bid) and poly[ADP-ribose]polymerase 1 (PARP-1), thereby inducing tumor cell death.

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Therapeutic Use
/Clinical Trials/ ClinicalTrials.gov is a registry and results database that includes publicly and privately funded human clinical studies worldwide. This website is maintained by the National Library of Medicine (NLM) and the National Institutes of Health (NIH). Each record on ClinicalTrials.gov provides a summary of the study protocol, including: the disease or condition; the intervention (e.g., the medical product, behavior, or procedure being investigated); the title, description, and design of the study; participation requirements (eligibility criteria); the location of the study; contact information for the study location; and links to relevant information from other health websites, such as NLM's MedlinePlus (for providing patient health information) and PubMed (for providing citations and abstracts of academic articles in the medical field). Toluenesulfonamide is included in the database.
/EXPL THER/ Severe malignant airway obstruction (SMAO) is a life-threatening type of non-small cell lung cancer (NSCLC). /The purpose of this study/ was to determine the efficacy and safety of intratumoral injection of toluenesulfonamide (PTS) for the treatment of NSCLC-SMAO. Ninety patients with non-small cell lung cancer with airway obstruction (NSCLC-SAO) received multiple intratumoral injections of PTS until the tumor volume shrank by 50% or more. The primary endpoint was objective response rate, assessed by chest computed tomography (CT) and bronchoscopy on days 7 and 30 after the last dose. Secondary endpoints included airway obstruction, lung function, quality of life, and survival. In the full analysis set (N=88), PTS treatment achieved a significant objective response rate according to RECIST criteria [day 7: chest CT: 59.1% (95% CI: 48.1%–69.5%), bronchoscopy: 48.9% (95% CI: 38.1%–59.8%); chest CT: 43.2% (95% CI: 32.7%–54.2%), bronchoscopy: 29.6% (95% CI: 20.3%–40.2%) (day 30). On day 7 post-treatment, FVC (mean difference: 0.35 L, 95% CI: 0.16–0.53 L), FEV1 (mean difference: 0.27 L, 95% CI: 0.07–0.48 L), baseline dyspnea index (mean difference: 64.8%, 95% CI: 53.9–74.7%), and the Cancer Treatment Functional Assessment – Lung Cancer subscale (mean difference: 6.9, 95% CI: 3.8–9.9) all significantly increased. We observed a significant decrease in the incidence of atelectasis (42.9%) and a significant decrease in the Eastern Cooperative Oncology Group (ECOG) performance status score (mean difference: 7.2, 95% CI: 3.9–10.5). The median survival for the full analysis set was 394 days, and the median survival for the protocol-compliant set was 460 days. Adverse events were reported in 64.0% of participants. Seven serious adverse events (7.9%) were reported, three of which resulted in death (one drug-related). Intratumoral injection of PTS was effective and well-tolerated as palliative treatment for non-small cell lung cancer (SMAO). The purpose of this study was to investigate the effect of percutaneous injection of p-toluenesulfonamide (PTS) on transplanted hepatocellular carcinoma in nude mice. Sixty nude mice with subcutaneous transplanted hepatocellular carcinoma were randomly divided into six groups: PTS group, chemotherapy group, radiotherapy group, PTS+chemotherapy group, PTS+radiotherapy group, and control group. PTS was injected into the nude mouse tumor model as instructed, and tumor growth rate and mouse survival time were recorded. All treatments effectively inhibited tumor growth, but the effects were more significant in the PTS+chemotherapy and PTS+radiotherapy groups. There was no significant difference in survival time among the groups. The PTS+chemotherapy and PTS+radiotherapy groups were safe and reliable, and their efficacy was superior to radiotherapy or chemotherapy alone. p-Toluenesulfonamide (PTS) has been shown to have anticancer effects against a variety of tumors. This study investigated the inhibitory effect of PTS on tongue squamous cell carcinoma (Tca-8113) and studied the changes in lysosomes and mitochondria after PTS treatment in vitro. High-performance liquid chromatography (HPLC) analysis showed that PTS selectively accumulated in Tca-8113 cells, while its concentration was relatively low in normal fibroblasts. Subsequently, we examined the effects of PTS on cell viability, invasion, and cell death. The results showed that PTS significantly inhibited the viability and invasiveness of Tca-8113 cells and increased cancer cell death. Flow cytometry analysis and lactate dehydrogenase release assays indicated that PTS induced cancer cell death by simultaneously activating apoptosis and necrosis. Observed morphological changes, such as cell shrinkage, nuclear condensation, and the formation of apoptotic bodies and secondary lysosomes, suggested that PTS may induce cell death by disrupting lysosomal stability. Lysosomal integrity assays and Western blotting experiments showed that PTS increased lysosomal membrane permeability, accompanied by activation of lysosomal cathepsin B. In addition, PTS also inhibited ATP biosynthesis and induced the release of mitochondrial cytochrome c. Therefore, our findings provide new insights into the application of PTS in cancer treatment. Hepatocellular carcinoma (HCC) is difficult to eradicate due to its refractory nature. Larger tumors often involve the portal vein, making surgical resection and local ablation therapies, such as percutaneous ethanol injection (PEI) and radiofrequency ablation (RFA), unsuitable for patients as these methods are considered neither effective nor safe. Currently, there are few effective treatments for HCC in this condition. Toluenesulfonamide (PTS) is a unique locally injectable lipophilic liquid antitumor drug with mild side effects, capable of rapidly and effectively necrotizing tumor tissue. Unlike PEI and RFA, PTS spreads more readily within the tumor than other corrosive agents such as alcohol. Therefore, PTS may offer additional benefits for patients with HCC involving vascular invasion. This article reports a case of a 70-year-old HCC patient who received PTS injection combined with transcatheter arterial chemoembolization (TACE) with significantly improved clinical outcomes.

C7H9NO2S

171.22

171.035

70-55-3

4-Tolyl-sulfonamide-d4;1219795-34-2

6269

Monoclinic plates (w+2)
White leaflets

1.3±0.1 g/cm3

322.2±35.0 °C at 760 mmHg

134-137 °C(lit.)

148.6±25.9 °C

0.0±0.7 mmHg at 25°C

1.564

0.79

1

3

1

11

209

0

S(C1C([H])=C([H])C(C([H])([H])[H])=C([H])C=1[H])(N([H])[H])(=O)=O

LMYRWZFENFIFIT-UHFFFAOYSA-N

InChI=1S/C7H9NO2S/c1-6-2-4-7(5-3-6)11(8,9)10/h2-5H,1H3,(H2,8,9,10)

4-methylbenzenesulfonamide

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.)