Triglycidyl isocyanurate (0–30 μM; 48 hours) suppresses the growth of human non-small-cell lung cancer spheroids in culture and causes the tumorspheres of A549, H460, and H1299 cells to gradually shrink[1]. Triglycidyl isocyanurate (0-30 μM; 48 hours) decreases the expression of Akt1/2/3 and phosphorylated Aktser473/474/472 of A549, H460, and H1299 tumorspheres; however, only A549 and H460 tumorspheres exhibit evident PARP and procaspase-3 cleavage along with the emergent active caspase-3 fragment[1]. Isocyanurate triglycidyl

Triglycidyl isocyanurate (0–30 μM; 48 hours) suppresses the growth of human non-small-cell lung cancer spheroids in culture and causes the tumorspheres of A549, H460, and H1299 cells to gradually shrink[1]. Triglycidyl isocyanurate (0-30 μM; 48 hours) decreases the expression of Akt1/2/3 and phosphorylated Aktser473/474/472 of A549, H460, and H1299 tumorspheres; however, only A549 and H460 tumorspheres exhibit evident PARP and procaspase-3 cleavage along with the emergent active caspase-3 fragment[1]. Isocyanurate triglycidyl

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Treatment with Teroxirone (5, 10, 30 µM for 48 h) dose-dependently reduced the size and formation of multicellular tumorspheres derived from human non-small cell lung cancer (NSCLC) cell lines H460, A549 (wild-type p53), and H1299 (p53-null) in suspension culture. [1]
In a soft agar colony formation assay, Teroxirone (10, 30, 50 µM) significantly diminished the number of colonies formed by tumorspheres from A549, H460, and H1299 cells after 30 days of incubation. The concentrations required to reduce colony numbers to less than 50% of control ranged between 30 and 50 µM. [1]
Western blot analysis of protein lysates from tumorspheres treated with Teroxirone (5, 10, 30 µM, 48 h) showed a dose-dependent reduction in the expression of stemness markers Nanog, ALDH1A1, and CD44 in H460 and A549 spheres. A similar reduction in Nanog was observed in H1299 spheres. [1]
Teroxirone treatment (5, 10, 30 µM, 48 h) induced cleavage of PARP and procaspase-3, and the appearance of active caspase-3 fragment in H460 and A549 tumorspheres (wild-type p53), accompanied by a dose-dependent increase in p53 protein levels. These apoptotic markers were not detected in p53-null H1299 tumorspheres. [1]
Teroxirone inhibited the expression of total Akt1/2/3 and phosphorylated Akt (Ser473/472/474) in tumorspheres from all three cell lines. [1]
BrdU incorporation assay showed that Teroxirone (5, 10, 30 µM, 48 h) dose-dependently reduced DNA synthesis/proliferation in tumorspheres from H460, A549, and H1299 cells. [1]
TUNEL assay confirmed that Teroxirone (5, 30 µM, 48 h) induced apoptotic cell death specifically in tumorspheres from H460 and A549 cells (wild-type p53), as indicated by increased TUNEL fluorescence. No significant TUNEL signal was observed in p53-null H1299 tumorspheres. [1]
Teroxirone inhibited the growth of human hepatocellular carcinoma (HCC) cell lines in a dose-dependent manner. The IC50 for Huh7 cells (mutant p53) was approximately 5 µM after 48 hours of treatment. No significant growth inhibition was observed in HepG2 (wild-type p53) and Hep3B (p53-null) HCC cells, or in normal hepatic LO2 cells within the tested concentration range (2-20 µM).[2]
Teroxirone induced apoptosis specifically in Huh7 cells, as evidenced by an increase in the sub-G1 population and Annexin V-FITC/PI staining. At 10 µM, it induced 6% early and 32% late apoptosis.[2]
Western blot analysis revealed that Teroxirone treatment (2-10 µM, 48h) in Huh7 cells led to cleavage/activation of caspase-8, -6, -3, and PARP, and increased levels of Fas and FADD. No changes were observed in intrinsic pathway markers like Bcl-2 and procaspase-9.[2]
At sub-apoptotic concentrations (0.5-2 µM), Teroxirone significantly inhibited the migration and invasion of Huh7 cells in transwell assays. It also inhibited wound healing in a scratch assay.[2]
Gelatin zymography and Western blot showed that Teroxirone (0.5-2 µM) reduced the secretion and activity of matrix metalloproteinases MMP-2 and MMP-9, as well as VEGF, in Huh7 cells.[2]
Pretreatment with the caspase-3 inhibitor z-DEVD-FMK (20 µM) reversed the cytotoxic effects, restored cell viability, and abrogated the cleavage of caspases and PARP induced by Teroxirone.[2]

In subcutaneous injection, tiglycidyl isocyanurate (1.8 and 3.6 mg/kg; every 2-3 days for a total of seven times; 30 days) inhibits the formation of xenograft tumors and has no effect on the weight of the mice in the nude[2].

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In a subcutaneous xenograft model using nude mice inoculated with Huh7 cells, administration of Teroxirone (1.8 and 3.6 mg/kg, subcutaneous, every 2-3 days for a total of 7 doses) significantly suppressed tumor growth compared to the PBS control group.[2]
Tumor weights were reduced in the Teroxirone-treated groups. No significant body weight loss was observed in treated mice.[2]
Histological examination (H&E staining) of resected tumors from Teroxirone-treated mice showed apoptotic features such as condensed cytoplasm and pyknosis.[2]
TUNEL assay confirmed increased apoptosis in tumors from Teroxirone-treated mice. Immunohistochemistry showed decreased levels of the proliferation marker PCNA and indicated activation of the extrinsic pathway markers Fas and FADD in treated tumors.[2]

Cell Viability Assay[1]
Cell Types: A549, H460 and H1299 cells
Tested Concentrations: 0 μM; 5 μM; 10 μM; 30 μM
Incubation Duration: 48 hrs (hours)
Experimental Results: Inhibited tumor cells growth in soft agar.

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Western Blot Analysis[1]
Cell Types: A549, H460 and H1299 cells
Tested Concentrations: 0 μM; 5 μM; 10 μM; 30 μM
Incubation Duration: 48 hrs (hours)
Experimental Results: Inhibited akt1/2/3 expression and p-aktser473/474/472 expression of A549, H460 and H1299 tumorspheres

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Tumorsphere Enrichment and Culture: Human lung adenocarcinoma cells (H460, A549, H1299) were cultured in serum-free DMEM-F12 medium supplemented with N-2 Plus Media Supplement, B-27 Supplement, 20 ng/mL EGF, and 10 ng/mL bFGF in ultra-low attachment plates to form and maintain floating spheroids. Spheres were expanded by mechanical dissociation and re-plating. [1]
Soft Agar Colony Formation Assay: Tumorspheres (pre-formed for 8 days to >50 µm diameter) were treated with various concentrations of Teroxirone for 48 h. Subsequently, the treated spheres were cultured in soft agar (0.5% agarose top layer, 0.3% agar bottom layer) for 30 days. Colonies were stained with 0.002% crystal violet, and colonies containing more than 50 cells were counted as positive. [1]
Western Blot Analysis: Tumorspheres (formed from 1x10^6 cells/well) treated with Teroxirone (5, 10, 30 µM, 48 h) were harvested and lysed. Protein concentrations were determined, and equal amounts of protein were resolved by SDS-PAGE, transferred to nitrocellulose membranes, and probed with specific primary antibodies (e.g., anti-p53, anti-PARP, anti-caspase-3, anti-Akt, anti-p-Akt, anti-Nanog, anti-CD44, anti-ALDH1A1, anti-GAPDH). Blots were incubated with HRP-conjugated secondary antibodies and visualized using enhanced chemiluminescence. [1]
BrdU Incorporation Assay: Tumorspheres treated with Teroxirone for 48 h were incubated with 10 µM BrdU for 1 h. Cells were fixed with a glycine/ethanol fixative, incubated with an anti-BrdU antibody, followed by an Alexa Fluor 488-conjugated secondary antibody. Fluorescence was visualized and quantified using fluorescence microscopy and image analysis software. [1]
TUNEL Assay: Tumorspheres treated with Teroxirone for 48 h were subjected to TUNEL staining to detect apoptotic DNA fragmentation. Stained slides were observed under a fluorescence microscope, and TUNEL-positive signals were quantified using image analysis software. [1]
Cell Viability Assay: Cells (Huh7, HepG2, Hep3B, LO2) were seeded in 96-well plates at 3x10³ cells/well. After attachment, cells were treated with various concentrations of Teroxirone in media containing 2% serum for 48 hours. Cell viability was assessed using the MTT assay. MTT solution (0.5 mg/mL) was added and incubated for 3 hours. Formazan crystals were dissolved in DMSO, and absorbance was measured at 570 nm. IC50 was calculated.[2]
Apoptosis Analysis by Flow Cytometry: For cell cycle analysis, treated cells were fixed in 70% ethanol, treated with RNase A and propidium iodide (PI), and analyzed by flow cytometry to determine the sub-G1 (apoptotic) population. For Annexin V/PI staining, trypsinized cells were resuspended in binding buffer and incubated with Annexin V-FITC and PI before flow cytometric analysis to distinguish early and late apoptotic cells.[2]
Western Blot Analysis: Cells (2x10⁶ per well) were treated with Teroxirone for 48 hours, lysed, and proteins were separated by SDS-PAGE. After transfer to nitrocellulose membranes, blots were blocked and incubated with primary antibodies against apoptosis-related proteins (e.g., caspases, PARP, Fas, FADD, Bcl-2, p53) and corresponding HRP-conjugated secondary antibodies. Proteins were visualized using an ECL detection system.[2]
Migration and Invasion Assay: For migration, 1x10⁴ Huh7 cells in serum-free medium were placed in the upper chamber of a transwell insert (8.0 µm pore membrane). Medium with 15% FBS was in the lower chamber as a chemoattractant. Cells were treated with Teroxirone or DMSO for 48 hours. Non-migrated cells on the upper side were removed, and cells that migrated to the underside were fixed, stained with crystal violet, and counted. For invasion, the transwell membrane was pre-coated with Matrigel before seeding cells and performing the same procedure.[2]
Wound Healing Assay: Huh7 cells were seeded in 12-well plates to form a confluent monolayer. A scratch was made with a pipette tip. After washing, cells were incubated with Teroxirone or DMSO. Wound closure was monitored at 0, 24, and 48 hours under a microscope, and the distance between wound edges was measured using image analysis software.[2]
Gelatin Zymography: Cells were cultured in media with 1% FBS and treated with Teroxirone. After 48 hours, conditioned media were collected, concentrated, and electrophoresed on SDS-polyacrylamide gels containing gelatin. Gels were rinsed, incubated in developing buffer overnight, and then stained with Coomassie Blue to visualize clear bands corresponding to MMP-2 and MMP-9 activity.[2]

Animal/Disease Models: Female nu/nu (nude) mice with Huh7 cells subcutaneously (sc) injected into the dorsal area[2]
Doses: 1.8 mg/kg and 3.6 mg/kg
Route of Administration: subcutaneous (sc)injection; every 2 –3 days for total seven times; 30 days
Experimental Results: Inhibited the growth of xenograft tumors.

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A subcutaneous xenograft tumor model was established in female nude mice (3-4 weeks old). Huh7 cells (1x10⁶) suspended in a 1:1 mixture of PBS and Matrigel were injected into the dorsal area. When tumors reached 50-100 mm³ (about 7 days post-inoculation), mice were randomly divided into groups. Teroxirone was dissolved and administered subcutaneously at doses of 1.8 and 3.6 mg/kg body weight. The control group received an equal volume of PBS. Administration occurred every 2-3 days for a total of seven doses. Tumor dimensions were measured regularly, and tumor volume was calculated. Mice were sacrificed 10 days after the final injection. Tumors were excised, weighed, and processed for histological and immunohistochemical analysis (H&E, TUNEL, PCNA, Fas, FADD staining).[2]

Absorption, Distribution and Excretion
In oral (gavage) studies in mice, at least 17% of the administered dose was absorbed within 24 hours. Blood analysis showed that the absorption rate of triglycidyl isocyanurate in aqueous solution was twice that of triglycidyl isocyanurate in sesame oil. Triglycidyl isocyanurate was distributed in the liver, stomach, and testes (only these three tissues were studied). The only available human data comes from clinical trials of α-triglycidyl isocyanurate (intravenous administration), which showed a systemic clearance of 5.7 L/min. Less than 1% of the administered dose was recovered in urine within 24 hours. In studies of oral (gavage) and intravenous [14C]α-triglycidyl isocyanurate in rabbits, approximately 30% and 60-70% of the radioactivity was recovered in urine within 24 hours, respectively.
Metabolism/Metabolites
In oral (gavage) studies in mice, plasma analysis showed that triglycidyl isocyanurate is hydrolyzed to diol diepoxide, diol epoxide, and fully hydrolyzed tridiol. No free triglycidyl isocyanurate was detected 8 hours after administration.
In in vitro studies, rapid hydrolysis of triglycidyl isocyanurate by epoxide hydrolases was observed in mouse liver preparations. Hydrolysis was also observed in rat liver preparations, but not in rat lung preparations. Microsomal epoxide hydrolases were found to have higher activity in human liver tissue from two kidney donors than in rat liver, using triglycidyl isocyanate as a substrate.
…The metabolism of TGIC involves the hydrolysis of epoxide groups to generate tridiol derivatives, a process promoted by hepatic epoxide hydrolases rather than pulmonary epoxide hydrolases. Non-enzymatic hydrolysis of epoxide groups occurs under low pH conditions. Other mechanisms of TGIC metabolism have not been investigated. TGIC and/or its metabolites are primarily excreted in the urine. Urinary metabolites of TGIC have not yet been identified.

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Biological Half-Life
In a study of intravenous administration of [(14)C]α-triglycidyl isocyanurate in rabbits, the half-life of triglycidyl isocyanurate in the blood was <5 minutes.
The only available human data currently comes from clinical trials of α-triglycidyl isocyanurate (intravenous administration), which showed that the average half-life of α-triglycidyl isocyanurate in the blood is approximately 1 minute…

Toxicity Summary
Identification and Uses: Triglycidyl isocyanurate (TGT) is a white solid. TGT is used as a crosslinking agent in polymer synthesis, as an additive in plastics, rubbers, and adhesives, as a curing agent in polyester powder coatings, as a protective coating for electronic devices, and as a topcoat ink and solder resist ink. Human Studies: Reported human health effects include contact dermatitis and respiratory anaphylaxis. It may also cause serious eye damage. TGT did not induce chromosomal aberrations in human lymphocytes at concentrations up to 2500 ng/mL. Only one aberration was reported at each of the higher concentrations of 5000 ng/mL and 10000 ng/mL. Human trials have been conducted in the clinical development of α-TGT as an antitumor drug. In these studies, α-TGT was administered intravenously to cancer patients at doses up to 900 mg/kg body weight using various dosing regimens. Toxic reactions included myelosuppression, nausea, and vomiting, as well as rare high-dose (>600 mg/kg body weight) alopecia and leukopenia. α-TGT has not been developed as an antitumor drug due to its severe local toxicity at the injection site (thrombophlebitis). Animal studies: Acute animal toxicity studies showed that TGT is toxic via oral and inhalation routes, but has low acute skin toxicity. TGT can cause severe eye irritation. It is a skin sensitizer, but not a skin irritant. Short-term repeated-dose studies showed that TGT can cause damage to the kidneys, lungs, stomach/duodenum, and sperm cells. In a subchronic toxicity/fertility study in rats, only a dose-dependent decrease in sperm count was observed when the dietary TGT concentration was up to 100 ppm. The chemical showed positive results in a series of in vitro genotoxicity studies (genetic mutation, unplanned DNA synthesis, sister chromatid exchange, and chromosomal aberration assays in bacterial and mammalian cells). In vivo, TGT has also been observed to have genotoxic effects on somatic cells (bone marrow cells) and testicular germ cells. Genotoxicity studies showed that inhaled TGT caused cytotoxicity and chromosomal aberrations in mouse spermatogonia. A 13-week dietary study in rats showed that the drug had no effect on male fertility.

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Toxicity Data
LC50 (Rat)> 650 mg/m3/4h

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Non-human Toxicity Values
LD50 Rats Oral 188-715 mg/kg Body Weight
LD50 Rats Skin >2000 mg/kg Body Weight

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The study reported that in animal xenotransplantation models, administration of teroxirone at doses of 1.8 and 3.6 mg/kg did not result in a significant decrease in mouse body weight, indicating that there was no significant systemic toxicity at these doses and administration regimens. [2]

[1]. Teroxirone motivates apoptotic death in tumorspheres of human lung cancer cells. Chem Biol Interact. 2018 Aug 1;291:137-143.

[2]. Teroxirone suppresses growth and motility of human hepatocellular carcinoma cells.Biomed Pharmacother. 2018 Mar;99:997-1008.

Tris(2,3-epoxypropyl)isocyanurate is a white crystalline solid. (NTP, 1992) Taroxidone is a triazine tricyclic oxide with antitumor activity. Taroxidone can alkylate and crosslink DNA, thereby inhibiting DNA replication. (NCI04) Taroxidone (1,3,5-triazine-2,4,6(1H,3H,5H)-trione-1,3,5-tri-(epoxymethyl)) is a synthetic tricyclic oxide derivative that has been studied as an anticancer drug. [1] Studies have shown that Taroxidone can effectively target tumor spherical carcinoma stem cells (CSCs) enriched from non-small cell lung cancer (NSCLC) cells. Its inhibitory effect on CSC proliferation is achieved by inducing p53-dependent apoptosis in cells expressing wild-type p53. [1]
Teroxirone can also reduce the expression of key stem cell markers (Nanog, ALDH1A1, CD44) and inhibit the Akt signaling pathway in tumor spheres. [1]
Results suggest that teroxirone may eradicate drug-resistant lung cancer stem cells by activating the tumor suppressor p53. [1]
Teroxirone (1,3,5-triazine-2,4,6(1H,3H,5H)-trione-1,3,5-tri-(ethylene oxide methyl)) is a low molecular weight tricyclic oxide derivative. Its mechanism of action is believed to be to bind to cancer cell nucleoproteins, causing irreparable DNA damage, which is quite different from platinum drugs that interact directly with DNA. [2]
This study shows that Teroxirone mainly exerts its anti-tumor effect by activating the extrinsic apoptosis pathway (Fas/FADD/caspase-8/-6/-3), targeting hepatocellular carcinoma cells expressing mutant p53 (Huh7), and inhibiting metastatic potential by inhibiting MMP-2, MMP-9, and VEGF. [2]
It has selective cytotoxicity against hepatocellular carcinoma cells expressing mutant p53 (Huh7), but is not toxic to cells expressing wild-type p53 (HepG2) or cells lacking p53 (Hep3B) or normal hepatocytes. [2]
This study suggests that Teroxirone may be a potential drug for the treatment of hepatocellular carcinoma, especially suitable for the treatment of tumors. Cells carrying p53 mutations are usually resistant to conventional chemotherapy. [2]

C12H15N3O6

297.26

297.096

2451-62-9

28825-96-9

17142

White to off-white solid powder

1.6±0.1 g/cm3

501.1±15.0 °C at 760 mmHg

95-98°C

256.9±20.4 °C

0.0±1.3 mmHg at 25°C

1.635

-2.77

0

6

6

21

416

0

OUPZKGBUJRBPGC-UHFFFAOYSA-N

InChI=1S/C12H15N3O6/c16-10-13(1-7-4-19-7)11(17)15(3-9-6-21-9)12(18)14(10)2-8-5-20-8/h7-9H,1-6H2

1,3,5-tris(oxiran-2-ylmethyl)-1,3,5-triazinane-2,4,6-trione

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)

DMSO : 50 mg/mL (168.20 mM)

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.)