DNA/RNA Synthesis

The most common types of bleomycin-induced DNA damage are single- and double-strand breaks and short-circuiting of individual desulfide junctions/desulfation switches. Bleomycin is a true radiomimetic compound that mimics the genetic effects of ionizing radiation [1]. The IC50 value of bleomycin hydrochloride against UT-SCC-19A cell line is 4.0±1.3 nM. Both UT-SCC Bleomycin Hydrochloride (50, 100 μM; 24, 48)-12A and UT-SCC-12B have strong resistance to bleomycin; the IC50 values are 14.2±2.8 nM and 13.0 respectively. ±1.1 nM[2]. h) Induces pulmonary fibrosis in RLE-6TN cells (50 μM) and A549 cells (100 μM) [4].

Bleomycin hydrochloride can be used in animal modeling to generate animal pulmonary fibrosis models. After treatment with bleomycin hydrochloride (3.5-4.0 mg/kg; intratracheal agent) on day 0, body weight decreased on day 4, followed by increased bleomycin hydrochloride (3.5-4.0 mg/kg; intratracheal agent) on day 7; Intratracheal preparation) can significantly increase pulmonary hypertensive concentrations and increase the right caudal lobe mass [3]. Instillation; 5.0 mg/kg/day) produced lung fibrosis in 80 8-week-old cosmetic BALB/c rats weighing approximately 20-30 g. Bleomycin induces α-SMA and collagen I expression levels [4] Bleomycin hydrochloride (intratracheal; 2.5 mg/kg; 1.25 mg/mL, roughly 50 μl per mouse) produces toy C57BL/6 model (8 weeks old, with an average weight of around 24.5 g) pulmonary fibrosis [5].

ADIPO-P2 cells are cultured in D-MEM high glucose medium at 37 °C with 5% CO₂ atmosphere, supplemented with 20% fetal calf serum, penicillin (100 U/mL), and streptomycin (100 μg/mL). 1.5 × 10⁵ cells/mL are cultured as monolayers in TC25 Corning flasks. Two flasks are set up for each experiment: one for the treated culture and one for the control. ADIPO-P2 cells are exposed to a 30-minute pulse of 2.5 μg/mL bleomycin sulfate during the log phase of growth. Parallel cultures serving as controls are not subjected to bleomycin sulfate. The duration and concentration of bleomycin sulfate exposure are selected based on earlier research using bleomycin sulfate exposure in mammalian cells conducted in our lab. The cells are maintained in culture with fresh culture medium until harvesting after being twice washed with Hank's balanced salt solution following the completion of the Bleomycin sulfate pulse treatment. After treatment, cells are kept in culture continuously for five passages or subcultures. When the cultures reach confluency (approximately 4 × 10⁵ cells/mL of culture medium), subcultivation is performed. At the time of subcultivation, cells are collected by trypsinization, and the number of viable cells is determined by staining an aliquot of approximately 200 μL with 0.4% trypan blue. This process allows for the estimation of cell growth. Subsequently, the cells are suspended in new culture medium and added to fresh culture flasks with a density of 1 × 10⁵5 cells/mL to continue growing. After the treatments are over, the remaining cells are either thrown away or transferred to another flask for cytogenetic analysis, which takes place 18 hours and 10 days later. Colchicine (0.1 μg/mL) is added to cell cultures in the final three hours of culture to analyze chromosomal aberrations. Standard protocols are followed when preparing chromosomes. Following harvesting, cells undergo hypotonic shock, are fixed in a 3:1 methanol:acetic acid solution, are spread out onto glass slides, and then undergo PNA-FISH processing. There are two separate experiments conducted.

Animal/Disease Models: Male Fischer 344 rats, 8-10 weeks old, body weight 150-250 g[3]
Doses: 3.5-4 mg/kg
Route of Administration: intratracheal
Experimental Results: Body weight diminished on the 4th day, increased on the 7th day, Ended by research.

Absorption, Distribution and Excretion
Systemic absorption is approximately 45%.
It was reported that patients with moderately severe renal failure excreted less than 20% of the dose in the urine.
Bleomycin sulfate is not significantly absorbed from the GI tract and the drug must be administered parenterally. Bleomycin is absorbed systemically following intrapleural or intraperitoneal administration. Systemic absorption of 45% has been reported following intrapleural administration of bleomycin.
Bleomycin is rapidly absorbed following either intramuscular (IM), subcutaneous (SC), intraperitoneal (IP) or intrapleural (IPL) administration reaching peak plasma concentrations in 30 to 60 minutes. Systemic bioavailability of bleomycin is 100% and 70% following IM and SC administrations, respectively, and 45% following both IP and IPL administrations, compared to intravenous and bolus administration.
Bleomycin is widely distributed throughout the body with a mean volume of distribution of 17.5 L/sq m in patients following a 15 units/sq m IV bolus dose.
Protein binding of bleomycin is very low (1%).
For more Absorption, Distribution and Excretion (Complete) data for BLEOMYCIN (9 total), please visit the HSDB record page.

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Metabolism / Metabolites
Hepatic
Biotransformation is unknow; probably by enzymatic degradation in tissues (based on animal studdies). Tissue enzyme activity varies, which may determine toxicity and antitumor effect of bleomycin... It is not known if any of the metabolites are active.
Bleomycin is inactivated by a cytosolic cysteine proteinase enzyme, bleomycin hydrolase. The enzyme is widely distributed in normal tissues with the exception of the skin and lungs, both targets of bleomycin toxicity. Systemic elimination of the drug by enzymatic degradation is probably only important in patients with severely compromised renal function.

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Biological Half-Life
115 minutes
In patients with creatinine clearance exceeding 35 mL/minute, the serum or plasma terminal half-life of bleomycin is about 2 hours. In patients with creatinine clearances less than 35 mL/minute, the terminal half-life of the drug is inversely related to creatinine clearance.
The average steady-state concentration of bleomycin in plasma of patients receiving continuous infusions of 30 units daily for 4-5 days is approx 150 ng/mL, and there is little bound to plasma proteins. Bleomycin disappears from plasma in a biphasic fashion; the initial half-life is about 1.3 hr, & the terminal half-life is approximately 9 hr.

Interactions
General anesthetic use in patients previously treated with bleomycin may result in rapid pulmonary deterioration because bleomycin causes sensitization of lung tissue to oxygen; even with concentrations of inspired oxygen considered to be safe, pulmonary fibrosis may develop postoperatively.
Concurrent use of /antineoplastics or radiation therapy/ may result in increased bleomycin toxicity, including bone marrow depression, which is rarely caused by bleomycin alone, and mucosal and pulmonary toxicity...
Cisplatin-induced renal function impairment may result in delayed clearance and bleomycin toxicity even at low doses; caution is recommended because of the frequent combined use of these two agents.
Raynaud's phenomenon has occurred in patients receiving bleomycin and vinblastine, with or without cisplatin, and in a few patients receiving bleomycin as a single agent. Cisplatin-induced hypomagnesemia may be an additional, although not essential, factor associated with its occurrence in patients receiving combination regimens including bleomycin and cisplatin. The cause of Raynaud's phenomenon in these cases, however, is not clearly established and may involve the underlying disease or vascular compromise, bleomycin, vinblastine, hypomagnesemia, or some combination of these factors.
During chemotherapy with bleomycin and etoposide a 28-year-old male, suffering from germ cell cancer, developed acute myocardial infarction. Under treatment with heparin and aspirin the patient revealed no Q-waves in ECG and recovery was without complications. Four weeks after onset of infarction, thallium-201 scintigraphy showed only a small irreversible, posteroseptal perfusion defect; coronary angiography was not performed. The chemotherapy regimen was continued and modified to etoposide as well as cisplatin and ifosfamide without recurrence of cardiac symptoms or ECG changes.

[1]. Comparative analysis of individual chromosome involvement in micronuclei induced by bleomycin in human leukocytes. Mol Cytogenet. 2016 Jun 21;9:49.

[2]. Squamous cell cancer cell lines: sensitivity to bleomycin and suitability for animal xenograft studies. Acta Otolaryngol Suppl. 1997;529:241-4.

[3]. Therapeutic administration of inhaled INS1009, a LRX-15 prodrug formulation, inhibits bleomycin-induced pulmonary fibrosis in rats. Pulm Pharmacol Ther. 2018 Apr;49:95-103.

[4]. Scutellarein inhibits BLM-mediated pulmonary fibrosis by affecting fibroblast differentiation, proliferation, and apoptosis. Ther Adv Chronic Dis. 2020 Jul 30;11:2040622320940185.

[5]. Scutellarin ameliorates pulmonary fibrosis through inhibiting NF-κB/NLRP3-mediated epithelial-mesenchymal transition and inflammation. Cell Death Dis. 2020 Nov 13;11(11):978.

Therapeutic Uses
Antibiotics, Antineoplastic; Antibiotics, Glycopeptide; Antimetabolites, Antineoplastic
Bleomycin is indicated in the treatment of squamous cell carcinomas of the head and neck (including the mouth, tongue, tonsil, nasopharynx, oropharynx, sinus, palate, lip, buccal mucosa, gingiva, epiglottis, and larynx and paralarynx), cervix, penis, skin and vulva. It is also indicated for treatment of testicular carcinoma (including embryonal cell carcinoma, choriocarcinoma, and teratocarcinoma), esophageal, and thyroid carcinomas. /Included in US product label/
Bleomycin is indicated for the treatment of Hodgkin's and non-Hodgkin's lymphomas. /Included in US product label/
Bleomycin is indicated in the treatment of AIDS-associated Kaposi's sarcoma. /NOT included in US product label/
For more Therapeutic Uses (Complete) data for BLEOMYCIN (12 total), please visit the HSDB record page.

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Drug Warnings
The most serious toxic effect of bleomycin is pulmonary reactions, usually presenting as interstitial pneumonitis, which occurs in approximately 10% of patients receiving the drug. Bleomycin pneumonitis occasionally progresses to pulmonary fibrosis and has resulted in death in approximately 1% of patients receiving the drug. Pulmonary toxicity generally appears to be dose and age related, occurring most frequently in patients older than 70 years of age and those receiving a total dosage of more than 400 units; however, pulmonary toxicity is unpredictable and reportedly has developed in younger patients receiving lower doses (e.g. after a total dosage of less than 200 units). Fatal pulmonary fibrosis occurred in a geriatric patient who received a total dosage of only 20 units of the drug.
Rarely, sudden onset of an acute chest pain syndrome suggestive of pleuropericarditis has been reported during continuous infusions of bleomycin. Improvement of this syndrome may be noted with slower infusion rates of the drug and patients may require analgesics for treatment of pain; total recovery usually occurs after discontinuance of the drug.
In at least one patient, cavitary pulmonary nodules associated with granuloma developed after combination therapy containing bleomycin; spontaneous resolution of these lesions occurred despite continuation of therapy.
Careful monitoring for clinical manifestations and evidence of pulmonary toxicity is required in patients receiving bleomycin. Dosage modification or discontinuance of the drug may be necessary in patients experiencing pulmonary toxicity.
For more Drug Warnings (Complete) data for BLEOMYCIN (28 total), please visit the HSDB record page.

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Pharmacodynamics
Bleomycin is an antibiotic which has been shown to have antitumor activity. Bleomycin 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. Bleomycin 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. The antibiotic antitumor drugs are cell cycle-nonspecific except for Bleomycin (which has major effects in G2 and M phases).

C55H84CLN17O21S3

1451.00476741791

1449.487

67763-87-5

Bleomycin sulfate;9041-93-4

456190

White to light yellow solid powder

-7.5

20

31

36

96

2580

18

CC1=C(N=C(N=C1N)[C@H](CC(=O)N)NC[C@@H](C(=O)N)N)C(=O)N[C@@H](C(C2=CN=CN2)O[C@H]3[C@H]([C@H]([C@@H]([C@@H](O3)CO)O)O)O[C@@H]4[C@H]([C@H]([C@@H]([C@H](O4)CO)O)OC(=O)N)O)C(=O)N[C@H](C)[C@H]([C@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCCC5=NC(=CS5)C6=NC(=CS6)C(=O)NCCC[S+](C)C)O

OYVAGSVQBOHSSS-QRQYLRPSSA-O

InChI=1S/C55H83N17O21S3/c1-20-33(69-46(72-44(20)58)25(12-31(57)76)64-13-24(56)45(59)82)50(86)71-35(41(26-14-61-19-65-26)91-54-43(39(80)37(78)29(15-73)90-54)92-53-40(81)42(93-55(60)88)38(79)30(16-74)89-53)51(87)66-22(3)36(77)21(2)47(83)70-34(23(4)75)49(85)63-10-8-32-67-28(18-94-32)52-68-27(17-95-52)48(84)62-9-7-11-96(5)6/h14,17-19,21-25,29-30,34-43,53-54,64,73-75,77-81H,7-13,15-16,56H2,1-6H3,(H13-,57,58,59,60,61,62,63,65,66,69,70,71,72,76,82,83,84,85,86,87,88)/p+1/t21-,22+,23+,24-,25-,29-,30+,34-,35-,36-,37+,38+,39-,40-,41?,42-,43-,53+,54-/m0/s1

3-[[2-[2-[2-[[(2S,3R)-2-[[(2S,3S,4R)-4-[[(2S)-2-[[6-amino-2-[(1S)-3-amino-1-[[(2S)-2,3-diamino-3-oxopropyl]amino]-3-oxopropyl]-5-methylpyrimidine-4-carbonyl]amino]-3-[(2R,3S,4S,5S,6S)-3-[(2R,3S,4S,5R,6R)-4-carbamoyloxy-3,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-3-(1H-imidazol-5-yl)propanoyl]amino]-3-hydroxy-2-methylpentanoyl]amino]-3-hydroxybutanoyl]amino]ethyl]-1,3-thiazol-4-yl]-1,3-thiazole-4-carbonyl]amino]propyl-dimethylsulfanium

Bleomycin hydrochloride; 67763-87-5; DTXSID2042690; SCHEMBL21331830;

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)

H2O : ~100 mg/mL
DMSO : ~50 mg/mL

Solubility in Formulation 1: ≥ 2.5 mg/mL (Infinity 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 (Infinity 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: ≥ 2.5 mg/mL (Infinity mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly.

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Solubility in Formulation 4: 100 mg/mL (Infinity 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.)