PKC ( IC50 = 8.5 μM ); Topoisomerase II

To determine whether mitoxantrone (MXN) was effective against CPXV in vivo, 0.25 or 0.5 mg/kg of drug was administered intraperitoneally to 4–6 week old female C57Bl/6 mice 1 day post-intraperitoneal inoculation. Log-rank tests were performed comparing survival times between the two MXN treatment groups relative the mock-treated group. A p-value of 0.025 was considered statistically significant, after a Bonferroni correction for the comparisons of the two MXN dose groups to the mock-treated. The lower dose of MXN significantly improved the survival time of infected animals, with the median day-of-death (MDD) in infected animals increasing from day 7 to day 9 (p = 0.0015). The 0.5 mg/kg dose of MXN improved the MDD to 12 days (p = 0.0005). Twenty-five percent of the animals treated with 0.5 mg/kg MXN survived compared to 5% of mock-treated animals (Fig. 2 ). Administration of higher levels of MXN or additional dosages were not beneficial (data not shown).[1]

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Mitoxantrone administered at the ideal dose (1.6 mg/kg/day; as a free base) results in a statistically significant number of 60-day survivors (curative effect) in mice with IP implanted L1210 leukemia. Mitoxantrone and ADM given intravenously (IV) also demonstrate efficacious antitumor activities in Lewis lung carcinoma implanted in SC, producing a 60% and a 45% ILS, respectively.

Mitoxantrone inhibits PKC in a non-competitive manner with respect to phosphatidylserine and ATP, but in a competitive manner with respect to histone H1, where its Ki value is 6.3 μM. Cell viability is reduced when B-CLL cells are treated with mitoxantrone (0.5 μg/mL) for 48 hours. Poly(ADP-ribose) polymerase (PARP) is subjected to proteolytic cleavage and DNA fragmentation upon induction by mitoxantrone, indicating that the cytotoxic effect of the drug is a result of apoptosis induction. Human breast carcinoma cell lines MDA-MB-231 and MCF-7 exhibit cytotoxicity to mitoxantrone, with IC50 values of 18 and 196 nM, respectively.

In standard 96-well plates, the human breast carcinoma cell lines MDA-MB-231 and MCF-7 are seeded. The culture medium is swapped out for one containing varying concentrations of mitoxantrone (10-5 to 5 μM) with or without DHA (30 μM) for a period of seven days following seeding. The tetrazolium salt assay is used to determine the overall viability of cells.

In agreement with a previous report which found that mitoxantrone/MXN exhibited no in vivo activity against intranasal VACV infection in BALB/c mice (Deng et al., 2007), we observed no efficacy against intranasal CPXV infection in BALB/c mice. Mitoxantrone did, however, demonstrate efficacy when used to treat intraperitoneally-infected C57Bl/6 mice, suggesting that differences in the route of infection and in mouse strain susceptibility to infection may influence MXN’s efficacy. While related anthracenediones have been reported to have in vivo antiviral activity against viruses unrelated to poxviruses (Dang et al., 2009, Sill et al., 1974), to our knowledge this is the first report of limited in vivo antiviral activity by MXN against poxviruses.[1]

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1.6 mg/kg/day; i.p. or i.v.

Mice

Absorption, Distribution and Excretion
Poorly absorbed following oral administration
1000 L/m2
21.3 L/hr/m2 [Elderly patients with breast cancer receiving IV administration of 15-90 mg/m2]
28.3 L/hr/m2 [Non-elderly patients with nasopharyngeal carcinoma receiving IV administration of 15-90 mg/m2]
16.2 L/hr/m2 [Non-elderly patients with malignant lymphoma receiving IV administration of 15-90 mg/m2]

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Metabolism / Metabolites
Hepatic
Hepatic
Half Life: 75 hours

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Biological Half-Life
75 hours

Toxicity Summary
Mitoxantrone, a DNA-reactive agent that intercalates into deoxyribonucleic acid (DNA) through hydrogen bonding, causes crosslinks and strand breaks. Mitoxantrone also interferes with ribonucleic acid (RNA) and is a potent inhibitor of topoisomerase II, an enzyme responsible for uncoiling and repairing damaged DNA. It has a cytocidal effect on both proliferating and nonproliferating cultured human cells, suggesting lack of cell cycle phase specificity.

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Hepatotoxicity
Chemotherapy with mitoxantrone alone is associated with serum enzyme elevations in up to 40% of patients, but these elevations are generally mild-to-moderate in severity, transient and not accompanied by symptoms or jaundice. Higher rates of liver enzyme elevations have been reported with combination chemotherapeutic regimens that include mitoxantrone. In high doses, mitoxantrone has been associated with a high rate of jaundice, but the degree of hyperbilirubinemia has been mild, transient and not associated with significant serum enzyme elevations or evidence of hepatitis. Rare instances of acute liver injury have been reported in patients taking mitoxantrone, including a single case of drug-rash with eosinophilia and systemic symptoms (DRESS). The latency to onset was 8 weeks and the pattern of serum enzyme elevations was cholestatic and later mixed. Immunoallergic features were prominent and appeared to respond to corticosteroid therapy. Other drugs were being taken and the association with mitoxantrone was not definite (Case 1). Thus, idiosyncratic and clinically apparent liver injury from mitoxantrone may occur but is quite rare.
Likelihood score: D (possible rare cause of clinically apparent liver injury).

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Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
Most sources consider breastfeeding to be contraindicated during maternal antineoplastic drug therapy, such as mitoxantrone. It might be possible to breastfeed safely during intermittent therapy with an appropriate period of breastfeeding abstinence, but the duration of abstinence is not clear. In one patient, mitoxantrone was still detectable in milk 28 days after a dose of 6 mg per square meter. Chemotherapy may adversely affect the normal microbiome and chemical makeup of breastmilk. Women who receive chemotherapy during pregnancy are more likely to have difficulty nursing their infant.
◉ Effects in Breastfed Infants
One mother received 3 daily doses of 6 mg/sq. m. of mitoxantrone intravenously along with 5 daily doses of etoposide 80 mg/sq. m. and cytarabine 170 mg/sq. m. intravenously. She resumed breastfeeding her infant 3 weeks after the third dose of mitoxantrone at a time when mitoxantrone was still detectable in milk. The infant had no apparent abnormalities at 16 months of age.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

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Protein Binding
78%

[1]. Antiviral Res. 2011 Dec 11;93(2):305–308. Inhibition of cowpox virus and monkeypox virus infection by mitoxantrone

[2]. Mechanism of action of mitoxantrone. Neurology.2004 Dec 28;63(12 Suppl 6):S15-8.

Pharmacodynamics
Mitoxantrone 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.

C22H29CLN4O6.HCL

517.4

516.154

C, 51.07; H, 5.84; Cl, 13.70; N, 10.83; O, 18.55

70476-82-3

65271-80-9; 70476-82-3 (HCl salt); 70711-41-0 (diacetate)

4212

Black solid powder

805.7ºC at 760 mmHg

203-205ºC

441.1ºC

2.392

8

10

12

32

571

0

Cl[H].Cl[H].O=C1C2=C(C([H])=C([H])C(=C2C(C2=C(C([H])=C([H])C(=C21)N([H])C([H])([H])C([H])([H])N([H])C([H])([H])C([H])([H])O[H])N([H])C([H])([H])C([H])([H])N([H])C([H])([H])C([H])([H])O[H])=O)O[H])O[H]

ZAHQPTJLOCWVPG-UHFFFAOYSA-N

InChI=1S/C22H28N4O6.2ClH/c27-11-9-23-5-7-25-13-1-2-14(26-8-6-24-10-12-28)18-17(13)21(31)19-15(29)3-4-16(30)20(19)22(18)32;;/h1-4,23-30H,5-12H2;2*1H

1,4-dihydroxy-5,8-bis[2-(2-hydroxyethylamino)ethylamino]anthracene-9,10-dione;dihydrochloride

NSC-301739; CL-232325; NSC301739; CL 232325; NSC 301739; DHAQ; CL232325; Mitozantrone; Mitoxantrone HCl; Mitoxantrone dihydrchloride; US brand name: Novantrone. NSC 301739; DHAQ; CL232325; Foreign brand names: Mitroxone; Neotalem; Onkotrone; Pralifan

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.

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: ≥ 2.5 mg/mL (4.83 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 25.0 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: ≥ 2.5 mg/mL (4.83 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 25.0 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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Solubility in Formulation 3: Saline: 30 mg/mL

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Solubility in Formulation 4: 2 mg/mL (3.87 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication (<60°C).

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