Traditional Cytotoxic Agents

Mitomycin C (400 μM) is the clinical first choice for superficial bladder tumors because it significantly inhibits intravesical tumor growth in a rat bladder tumor model.[4]

DNA interstrand crosslinks (ICLs) are the most toxic lesions induced by chemotherapeutic agents such as mitomycin C and cisplatin. By covalently linking both DNA strands, ICLs prevent DNA melting, transcription, and replication. Studies on ICL signaling and repair have been limited, because these drugs generate additional DNA lesions that trigger checkpoint signaling. Here, we monitor sensing, signaling from, and repairing of a single site-specific ICL in cell-free extract derived from Xenopus eggs and in mammalian cells. Notably, we demonstrate that ICLs trigger a checkpoint response independently of origin-initiated DNA replication and uncoupling of DNA polymerase and DNA helicase. The Fanconi anemia pathway acts upstream of RPA-ATR-Chk1 to generate the ICL signal. The system also repairs ICLs in a reaction that involves extensive, error-free DNA synthesis. Repair occurs by both origin-dependent and origin-independent mechanisms. Our data suggest that cell sensitivity to crosslinking agents results from both checkpoint and DNA repair defects.[1]

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The discovery of the molecular targets of chemotherapeutic medicines and their chemical footprints can validate and improve the use of such medicines. In the present report, we investigated the effect of mitomycin C (MMC), a classical chemotherapeutic agent on cancer cell apoptosis induced by TRAIL. We found that MMC not only potentiated TRAIL-induced apoptosis in HCT116 (p53-/-) colon cancer cells but also sensitized TRAIL-resistant colon cancer cells HT-29 to the cytokine both in vitro and in vivo. MMC also augmented the pro-apoptotic effects of two TRAIL receptor agonist antibodies, mapatumumab and lexatumumab. At a mechanistic level, MMC downregulated cell survival proteins, including Bcl2, Mcl-1 and Bcl-XL, and upregulated pro-apoptotic proteins including Bax, Bim and the cell surface expression of TRAIL death receptors DR4 and DR5. Gene silencing of DR5 by short hairpin RNA reduced the apoptosis induced by combination treatment of MMC and TRAIL. Induction of DR4 and DR5 was independent of p53, Bax and Bim but was dependent on c-Jun N terminal kinase (JNK) as JNK pharmacological inhibition and siRNA abolished the induction of the TRAIL receptors by MMC[2].

Human colon cancer cells, HT-29 and colon adenocarcinoma HCT116, are employed respectively. The number of viable cells present in the culture is indicated by the luminescent signal generated by the CellTiter-Glo Luminescent Cell Viability Assay, which measures ATP using a special, stable form of luciferase. After exposing the cells to varying concentrations of TRAIL for 12 hours, the cells are pretreated with 5 μM of mitomycin C for either 12 or 24 hours. After adding an equivalent volume (100 μL) of CellTiter-Glo^TM reagent, the mixture is carefully mixed on an orbital shaker for two minutes. After allowing the luminescent signal to stabilize for ten minutes at room temperature, the mixture is imaged using the Xenogen IVIS system to determine the viability of the cells.

Absorption, Distribution and Excretion
Erratic.
Approximately 10% of a dose of mitomycin is excreted unchanged in the urine.
FOLLOWING IV INJECTION OF 2 MG/KG BODY WT ... WISTAR RATS, 18% WAS RECOVERED UNCHANGED IN URINE WITHIN 24 HR AT ... 8 MG/KG ... 35% WAS RECOVERED IN URINE, BUT NONE IN FECES OR TISSUES.
THIRTY MIN AFTER IV INJECTION OF 8 MG/KG BODY WT TO MICE TRACES REMAINED IN BLOOD. IN GUINEA PIGS DRUG WAS CONCN IN KIDNEYS & NOT IN LIVER, SPLEEN OR BRAIN & WAS EXCRETED IN URINE.
Mitomycin is absorbed inconsistently from the gastrointestinal tract, and it is therefore administered intravenously. It disappears rapidly from the blood after injection. Peak concentrations in plasma are 0.4 ug/ml after doses of 20 mg/m sq ... The drug is widely distributed throughout the body but is not detected in the brain.
In animals, highest mitomycin concentrations are found in the kidneys, followed by muscles, eyes, lung, intestines, and stomach. The drug is not detectable in the liver, spleen, or brain which rapidly inactivate mitomycin. Higher concentrations of the drug are generally present in cancer tissues than in normal tissues.
For more Absorption, Distribution and Excretion (Complete) data for MITOMYCIN C (9 total), please visit the HSDB record page.

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Metabolism / Metabolites
Primarily hepatic, some in various other tissues.
SUGGESTED ALKYLATING METABOLITES OF CARCINOGENS: MITOMYCIN C: REDUCTION PRODUCTS. /FROM TABLE/
Inactivation occurs by metabolism, but the products have not been identified. It is metabolized primarily in the liver, and less than 10% of the active drug is excreted in the urine or the bile.
The drug is eliminated primarily by hepatic metabolism with about 20% hepatic extraction and 10-30% recovery of intact drug in the urine. Clearance is 0.3-0.4 l/hr/kg.
Mitomycin disappears rapidly from the blood after intravenous injection. It is widely distributed but does not appear to cross the blood-brain barrier. Mitomycin is metabolized mainly in the liver; up to 10% of a dose is excreted unchanged in the urine.
MITOMYCIN C WAS PREFERENTIALLY ACTIVATED & METABOLIZED BY SONICATED CELL PREPARATIONS. BIOACTIVATION OF MITOMYCIN TO ALKYLATING AGENT BY EMT6 & SARCOMA 180 CELL SONICATES REQUIRED HYPOXIC CONDITIONS & NADPH-GENERATING SYSTEM.
Primarily hepatic, some in various other tissues.
Route of Elimination: Approximately 10% of a dose of mitomycin is excreted unchanged in the urine.
Half Life: 8-48 min

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Biological Half-Life
8-48 min
After doses of 20 mg/m sq ... Mitomycin is cleared from plasma with a half-time of approximately 1 hour.
/Mitomycin/ has an alpha half-life of 5-10 min after IV injection and beta half-life of 46 min.

Toxicity Summary
Mitomycin is activated in vivo to a bifunctional and trifunctional alkylating agent. Binding to DNA leads to cross-linking and inhibition of DNA synthesis and function. Mitomycin is cell cycle phase-nonspecific.

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Hepatotoxicity
Chemotherapy with mitomycin in combination with other agents is associated with serum enzyme elevations in a proportion of patients, depending upon the dose and other agents used. ALT elevations during mitomycin therapy are usually asymptomatic and transient and may resolve without dose modification. In many instances, it is difficult to attribute the liver test abnormalities to mitomycin, because of the exposure to other potentially hepatotoxic agents. High doses of mitomycin have been linked to cases of sinusoidal obstruction syndrome, typically presenting with right upper quadrant pain 10 to 30 days after the infusion, followed by weight gain, ascites and liver test abnormalities. Fatalities due to hepatic failure have occurred, but most patients recover within 1 to 3 months of onset. The frequency of sinusoidal obstruction syndrome limits the dosage of mitomycin that can be used in cancer chemotherapy and in myeloablation in preparation for bone marrow transplantation. There have been no convincing instances of acute, clinically apparent idiosyncratic liver injury with jaundice associated with mitomycin therapy.
Likelihood score: B[H] (very likely but now uncommon cause of sinusoid obstruction syndrome when given in high doses and in combination with other cytotoxic agents).

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Toxicity Data
LD50: 23 mg/kg (Oral, Mouse) (A308)
LD50: 30 mg/kg (Oral, Rat) (A308)

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Interactions
IN RATS GIVEN SINGLE DOSES OF 3 MG METHYLCHOLANTHRENE BY SC INJECTION INCIDENCE OF LOCAL SARCOMAS AFTER 120 DAYS WAS REDUCED WHEN WEEKLY IP INJECTIONS OF MITOMYCIN C WERE ALSO GIVEN.
IN MICE ... ADMIN 0.2 ML OF 1% SOLN OF METHYLCHOLANTHRENE IN BENZENE ON SKIN DAILY FOR 5-10 DAYS, INCIDENCE OF SKIN PAPILLOMAS WAS GREATLY INCR WHEN MITOMYCIN C WAS GIVEN DAILY BY 20 IP INJECTIONS ... .
IN RATS GIVEN 40 UG/KG BODY WT MITOMYCIN C IP & ORAL DOSE DMBA, INCIDENCE OF MAMMARY TUMORS AFTER 120 DAYS WAS SIMILAR TO THAT IN RATS GIVEN DMBA ALONE.
Absorption of cephalexin, sulfanilamide, salicylic acid, and D- and L-tryptophan was significantly decreased by the pretreatment with /iv/ mitomycin C /in rats/. Absorption of 6-carboxyfluorescein and fluorescein isothiocyanate conjugated dextran was not significantly affected by mitomycin C pretreatment. Maximal effects, using sulfanilamide as a model, were noted 48 hours after mitomycin C pretreatment. The dosage of mitomycin C ... did not affect the percentage of sulfanilamide absorbed.
For more Interactions (Complete) data for MITOMYCIN C (25 total), please visit the HSDB record page.

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Non-Human Toxicity Values
LD50 Mouse iv 5 mg/kg
LD50 cat iv 1-2.5 mg/kg
LD50 dog iv 1-2.5 mg/kg
LD50 monkey iv 1-2.5 mg/kg

[1]. Mol Cell . 2009 Sep 11;35(5):704-15.

[2]. Cell Cycle . 2012 Sep 1;11(17):3312-23.

[3]. Indian J Biochem Biophys . 2014 Feb;51(1):46-51.

[4]. Int J Oncol . 2014 Jan;44(1):147-52.

Therapeutic Uses
Antibiotics, Antineoplastic; Nucleic Acid Synthesis Inhibitors
Mitomycin is useful for the palliative treatment of gastric adenocarcinoma, in conjunction with fluorouracil and doxorubicin. It has produced temporary beneficial effects in carcinomas of the cervix, colon, rectum, pancreas, breast, bladder, head and neck, and lung, and in melanoma. It has also shown activity against lymphomas and leukemia, particularly chronic granulocytic leukemia, but not in myeloma.
Thirty patients with advanced colorectal adenocarcinoma were treated by chemotherapy with an alternating regimen consisting of 5-fluorouracil mitomycin C and 5-fluorouracil dacarbazine at 3 wk intervals. ... The toxicity of this regimen was essentially digestive with 30% of grade 3 or 4 nausea and vomiting. In spite of the reported active and synergistic action of drug association in colorectal carcinoma, this treatment schedule is not better than 5-fluorouracil alone. Gastrointestinal toxicity was incr.
Forty-two patients with metastatic breast cancer refractory to first line therapies were treated with combination chemotherapy with mitomycin-C and vinblastine. ... The toxicity was acceptable with 20 episodes of moderate myelosuppression (58.8%) and 2 cases with congestive heart failure that responded to medical treatment.
For more Therapeutic Uses (Complete) data for MITOMYCIN C (19 total), please visit the HSDB record page.

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Drug Warnings
Mitomycin is contraindicated in patients with pre-existing myelosupression & anemia.
Because normal defense mechanisms may be suppressed by mitomycin therapy, the patient's antibody response to the vaccine may be decreased. The interval between discontinuation of medications that cause immunosuppression and restoration of the patient's ability to respond to the vaccine depends on the intensity and type of immunosuppression-causing medication used, the underlying disease, and other factors; estimates vary from 3 months to 1 year.
cBecause normal defense mechanisms may be suppressed by mitomycin therapy, concurrent use with a live virus vaccine may potentiate the replication of the vaccine virus, may increase the side/adverse effects of the vaccine virus, and/or may decrease the patient's antibody response to the vaccine; immunization of these patients should be undertaken only with extreme caution after careful review of the patient's hematologic status and only with the knowledge and consent of the physician managing the cytarabine therapy. The interval between discontinuation of medication that cause immunosuppression and restoration of the patient's ability to respond to the vaccine depends on the intensity and type of immunosuppression-causing medications used, the underlying disease, and other factors; estimates vary from 3 months to 1 year. Patients with leukemia in remission should not receive live virus vaccine until at least 3 months after their last chemotherapy. In addition, immunization with oral polio-virus vaccine should be postponed in persons in close contact with the patient, especially family members.
Gonadal suppression, resulting in amenorrhea or azoospermia, may occur in patients taking antineoplastic therapy, especially with the alkylating agents. In general, these effects appear to be related to dose and length of therapy and may be irreversible. Prediction of the degree of testicular or ovarian function impairment is complicated by the common use of combinations of several antineoplastics, which makes it difficult to assess the effects of individual agents.
For more Drug Warnings (Complete) data for MITOMYCIN C (6 total), please visit the HSDB record page.

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Pharmacodynamics
Mitomycin is one of the older chemotherapy drugs, which has been around and in use for decades. It is an antibiotic which has been shown to have antitumor activity. Mitomycin selectively inhibits the synthesis of deoxyribonucleic acid (DNA). The guanine and cytosine content correlates with the degree of mitomycin-induced cross-linking. At high concentrations of the drug, cellular RNA and protein synthesis are also suppressed. Mitomycin has been shown in vitro to inhibit B cell, T cell, and macrophage proliferation and impair antigen presentation, as well as the secretion of interferon gamma, TNFa, and IL-2.

C15H18N4O5

334.37

334.127

C, 53.89; H, 5.43; N, 16.76; O, 23.93

50-07-7

5746

Black solid powder

1.9±0.1 g/cm3

532.0±60.0 °C at 760 mmHg

360 °C

275.5±32.9 °C

0.0±3.2 mmHg at 25°C

1.828

-0.27

3

8

4

24

757

4

NC1=C(C(C2=C(C1=O)[C@@H](COC(N)=O)[C@]3(OC)N2C[C@H]4[C@@H]3N4)=O)C

NWIBSHFKIJFRCO-WUDYKRTCSA-N

InChI=1S/C15H18N4O5/c1-5-9(16)12(21)8-6(4-24-14(17)22)15(23-2)13-7(18-13)3-19(15)10(8)11(5)20/h6-7,13,18H,3-4,16H2,1-2H3,(H2,17,22)/t6-,7+,13+,15-/m1/s1

[(4S,6S,7R,8S)-11-amino-7-methoxy-12-methyl-10,13-dioxo-2,5-diazatetracyclo[7.4.0.02,7.04,6]trideca-1(9),11-dien-8-yl]methyl carbamate

Ametycine; mitomycine C; Mitomycin; 50-07-7; Ametycine; Mutamycin; Mitomycin-C; Mitocin-C; Ametycin; mitomycinX. US trade names: Mitozytrex; Mutamycin. Foreign brand names: Ametycine; MitocinC; Mitolem; MitoMedac; Mutamycine. Abbreviations: MITC; MITO; MITOC; MTC; NCIC04706

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

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

Solubility in Formulation 1: ≥ 2.08 mg/mL (6.22 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (6.22 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
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.

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