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
The systemic absorption rate is approximately 45%. Less than 20% of the dose is reported to be excreted in the urine of patients with moderate to severe renal failure. Bleomycin sulfate is almost entirely not absorbed from the gastrointestinal tract and must be administered via parenteral route. Bleomycin is systemically absorbed after intrapleural or intraperitoneal administration. The systemic absorption rate of bleomycin after intrapleural administration is reported to be 45%. Bleomycin is rapidly absorbed after intramuscular (IM), subcutaneous (SC), intraperitoneal (IP), or intrapleural (IPL) injection, reaching peak plasma concentrations within 30 to 60 minutes. The systemic bioavailability of bleomycin after IM and subcutaneous injection is 100% and 70%, respectively, while the systemic bioavailability after intraperitoneal and retroperitoneal injection is 45%, compared to intravenous and bolus administration. Bleomycin is widely distributed throughout the body; after an intravenous bolus of 15 units/m², the mean volume of distribution in patients is 17.5 liters/m². Bleomycin has very low protein binding (1%). For more complete data on the absorption, distribution, and excretion of bleomycin (9 items in total), please visit the HSDB record page. Metabolism/Metabolites Liver Biotransformation is unclear; it may occur via enzymatic degradation in tissues (based on animal studies). Tissue enzyme activity varies, which may determine the toxicity and antitumor effects of bleomycin… It is currently unclear whether its metabolites are active. Bleomycin is inactivated by the cytoplasmic cysteine protease bleomycin hydrolase. This enzyme is widely distributed in normal tissues, except for the skin and lungs, which are target organs for bleomycin toxicity. Systemic drug clearance via enzymatic degradation may only be important in patients with severely impaired renal function.

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Biological Half-Life
115 minutes
For patients with creatinine clearance exceeding 35 mL/min, the serum or plasma terminal half-life of bleomycin is approximately 2 hours. For patients with creatinine clearance below 35 mL/min, the terminal half-life is negatively correlated with creatinine clearance.
In patients receiving a continuous infusion of 30 units of bleomycin daily for 4–5 days, the mean steady-state concentration of bleomycin in plasma is approximately 150 ng/mL, with extremely low binding to plasma proteins. Bleomycin clearance in plasma is biphasic; the initial half-life is approximately 1.3 hours, and the terminal half-life is approximately 9 hours.

Interactions
General anesthesia may lead to rapid deterioration of lung function in patients previously treated with bleomycin, as bleomycin makes lung tissue sensitive to oxygen; postoperative pulmonary fibrosis may still occur even if the inhaled oxygen concentration is considered safe. Concomitant use of antineoplastic drugs or radiotherapy may increase the toxicity of bleomycin, including bone marrow suppression (bleomycin alone rarely causes bone marrow suppression) and mucosal and pulmonary toxicity… Cisplatin-induced renal impairment may lead to delayed bleomycin clearance, and even low doses may cause bleomycin toxicity; caution is advised when using these two drugs in combination, as they are frequently used together. Raynaud's phenomenon has occurred in patients treated with bleomycin and vinblastine (with or without cisplatin) and in a small number of patients treated with bleomycin alone. Cisplatin-induced hypomagnesemia may be another contributing factor, although not a necessary one, to the occurrence of Raynaud's phenomenon in patients receiving bleomycin and cisplatin combined with chemotherapy. However, the etiology of Raynaud's phenomenon in these cases is unclear and may be related to underlying disease or vascular damage, bleomycin, vinblastine, hypomagnesemia, or some combination of these factors. A 28-year-old male with germ cell carcinoma experienced an acute myocardial infarction during bleomycin and etoposide chemotherapy. After treatment with heparin and aspirin, the patient's electrocardiogram (ECG) showed no Q waves and recovered smoothly. Four weeks after the infarction, thallium-201 myocardial scintigraphy revealed only a small, irreversible posterior septal perfusion defect; coronary angiography was not performed. The chemotherapy regimen continued and was adjusted to etoposide, cisplatin, and ifosfamide. No recurrence of cardiac symptoms or ECG changes was observed.

[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, antitumor drugs; antibiotics, glycopeptides; antimetabolites, antitumor drugs
Bleomycin is indicated for the treatment of squamous cell carcinoma of the head and neck (including the oral cavity, tongue, tonsils, nasopharynx, oropharynx, sinuses, palate, lips, buccal mucosa, gingiva, epiglottis, larynx, and paralarynx), cervix, penis, skin, and vulva. It is also indicated for the treatment of testicular cancer (including embryonal cell carcinoma, choriocarcinoma, and teratoma), esophageal cancer, and thyroid cancer. /Included on US product label/
Bleomycin is indicated for the treatment of Hodgkin's lymphoma and non-Hodgkin's lymphoma. /Included on US product label/
Bleomycin is indicated for the treatment of HIV-associated Kaposi's sarcoma. /Not included on US product label/
For more complete data on the therapeutic uses of bleomycin (12 types), please visit the HSDB record page.
Drug Warnings
The most serious toxicity of bleomycin is a pulmonary reaction, usually manifesting as interstitial pneumonia, which occurs in approximately 10% of patients using the drug. Bleomycin-induced pneumonia can sometimes progress to pulmonary fibrosis and cause death in approximately 1% of patients using the drug. Pulmonary toxicity is generally dose- and age-related, most commonly occurring in patients over 70 years of age and those receiving a total dose exceeding 400 units; however, pulmonary toxicity is difficult to predict, and it has been reported that younger patients may experience pulmonary toxicity even with lower doses (e.g., a total dose below 200 units). One elderly patient developed fatal pulmonary fibrosis after receiving only 20 units of bleomycin.
Rarely, a sudden acute chest pain syndrome, suggestive of pleuritic pericarditis, has been reported during continuous bleomycin infusion. Reducing the infusion rate can improve this syndrome; patients may require analgesics for pain management; full recovery is usually achieved upon discontinuation of the drug. At least one patient developed cavitary pulmonary nodules associated with granulomas after receiving combination therapy containing bleomycin; these lesions resolved spontaneously despite continued treatment. Patients receiving bleomycin should be closely monitored for clinical manifestations and signs of pulmonary toxicity. For patients experiencing pulmonary toxicity, dose adjustment or discontinuation may be necessary. For more complete data on drug warnings for bleomycin (28 in total), please visit the HSDB record page.

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Pharmacodynamics
Bleomycin is an antibiotic that has been shown to have antitumor activity. Bleomycin selectively inhibits deoxyribonucleic acid (DNA) synthesis. The levels of guanine and cytosine are correlated with the degree of mitomycin-induced cross-linking. At high concentrations, cellular RNA and protein synthesis are also inhibited. In vitro studies have shown that bleomycin inhibits the proliferation of B cells, T cells, and macrophages, and impairs antigen presentation and the secretion of interferon-γ, TNF-α, and IL-2. Except for bleomycin (which mainly acts on the G2 and M phases), other antibiotic antitumor drugs do not have cell cycle specificity.

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.)