Glycopeptide antitumor antibiotic

In a dose-dependent manner, peplomycin (0-100 μM; 30 h) inhibits the human oral squamous carcinoma cell line SSCKN and SCCTF cells[3]. Inducing apoptosis, peplomycin (1 μM and 10 μM; 30 h) causes dose-dependent nuclear fragmentation and chromatin condensation in cells. By differentiating fibroblasts to alpha-SMA-positive MF, peplomycin (5 mg/mL; 7 d) causes pulmonary fibrosis that is distinct from saline-injected rats (N-Fib)[4].

Rat pulmonary fibroblasts are converted to myofibroblasts (MF) by peplomycin (5 mg/kg; once daily), and the lungs' peripheries next to the pleura show advanced fibrosis with a small number of alpha-smooth muscle actin (alpha-SMA)-positive MF, which have an abundance of microfilaments and cellular organelles ultrastructurally[4].

Cell Viability Assay[3]
Cell Types: SSCKN and SCCTF cells
Tested Concentrations: 0, 1, 5, 10, 50, and 100 μM
Incubation Duration: 30 hrs (hours)
Experimental Results: diminished SSCKN and SCCTF cells viability dose-dependently up to 50 μM, and resulted the viability of 38% and 30% that of the control group, respectively.

Animals and their treatment [4]
Eight male specific pathogen-free Fisher 344 rats (8 weeks old) were used and five animals received serial intraperitoneal injection of 5 mg/kg peplomycin (PLM) for 10 days. The remaining three rats were injected sterile saline as the control. peplomycin (PLM)-injected and control rats were killed at 1 day and 2 weeks after the last injection and the lungs were immediately dissected. The pulmonary lobes were fixed with 4% paraformaldehyde dissolved in ribonuclease (RNase)-free water. Thin sections were prepared from fixed tissues and routinely stained with haematoxylin-eosin or immunohistochemically stained.

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Isolation of lung fibroblasts [4]
The peplomycin (PLM)-injected and saline-injected rats were fed for 2 weeks after the last injections then killed. The lungs were extirpated and the resected tissues were minced to pieces about 1–2 mm3, then the minced tissues were trypsinized and separated cells were cultivated in Dulbecco’s minimum essential medium (DMEM) containing 10% foetal bovine serum (FBS), streptomycin (100 μg/mL) and penicillin (100 units/mL). The cells were incubated at 37 C in a humidified atmosphere of 5% CO2/95% air. After two or three passages, the proliferated fibroblasts were subjected to the following experiments.

6852373 Intraperitoneal injection of LDLo 100 mg/kg in mice, Antibiotics Journal, 32(36), 1979 [PMID:83987]
6852373 Intravenous injection of LD50 51 mg/kg in mice, Future Drugs, 13(519), 1988

[1]. Peplomycin. Cancer Treat Rev. 1984 Dec;11(4):303-5.

[2]. A case of peplomycin-induced scleroderma. Br J Dermatol. 2004 Jun;150(6):1213-4.

[3]. Peplomycin-induced apoptosis in oral squamous carcinoma cells depends on bleomycin sensitivity. Oral Oncol. 2001 Jun;37(4):379-85.

[4]. Peplomycin, a bleomycin derivative, induces myofibroblasts in pulmonary fibrosis. Int J Exp Pathol. 2001 Aug;82(4):231-41.

Peplomycin is a glycopeptide antibiotic. Peplomycin is a semi-synthetic analogue of bleomycin, a mixture of several basic glycopeptide antitumor antibiotics isolated from Streptomyces verticillus. Peplomycin forms a complex with iron, reducing molecular oxygen to superoxide anions and hydroxyl radicals, leading to single- and double-strand breaks in DNA. The drug appears to have stronger antitumor activity than bleomycin; however, its use is limited due to its pulmonary toxicity. (NCI04) An antitumor drug derived from bleomycin. Oral squamous cell carcinoma cell lines SSCKN and SCCTF are highly sensitive to bleomycin, while SCCTF cells are extremely insensitive to the drug. To determine whether anticancer drug resistance in oral squamous cell carcinoma cells is related to the degree of drug-induced apoptosis, we investigated the apoptosis-inducing effect of Peplomycin on these cells. SCCKN and SCCTF cells were cultured to a subconvergent state and then exposed to different concentrations of Peplomycin. Phase-contrast microscopy and WST-1 cell viability assays showed that pepromycin induced cytotoxicity in both SCCKN and SCCTF cells in a dose-dependent manner, reaching its maximum effect at concentrations of 1 μM and 10 μM, respectively. Hoechst 33342 staining revealed significant nuclear condensation and chromatin fragmentation in SCCKN cells treated with 1 μM pepromycin. However, no nuclear condensation or fragmentation was observed in SCCTF cells treated with 1 μM pepromycin. DNA ladder banding was observed in both cell lines after pepromycin treatment. The formation of DNA ladder bands induced in SCCKN and SCCTF cells was dose-dependent, reaching its maximum effect at concentrations of 5 μM and 50 μM, respectively. Bleomycin also induced DNA ladder banding in both SCCKN and SCCTF cells, but with different sensitivities. Mitomycin C induced DNA ladder banding in both SCCKN and SCCTF cells; however, the intensity of DNA ladder banding was almost identical in both cell lines. The results of this study indicate that pepromycin-induced apoptosis in oral squamous cell carcinoma lines depends on the sensitivity of these cells to bleomycin. [3]

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To analyze the mechanism by which the bleomycin derivative pepromycin (PLM) induces pulmonary fibrosis, we investigated the differentiation of rat lung fibroblasts into myofibroblasts (MF). In rats that received intraperitoneal injection of PLM (5 mg/kg/day), severe fibrosis was observed in the peripheral lung region adjacent to the pleura, with a small number of α-smooth muscle actin (α-SMA)-positive microfibrils (MF) visible. The ultrastructure of these microfibrils showed that they contained abundant microfilaments and organelles. In the fibrotic tissue, the expression of α-SMA mRNA was detected by in situ reverse transcription-polymerase chain reaction (RT-PCR). The expression level of α-SMA mRNA increased significantly after 2 weeks of PLM administration and then gradually decreased, although fibrosis was still progressing. Lung fibroblasts (N-Fib) were isolated from rat lungs injected with physiological saline and cultured in a medium containing 5 mg/mL PLM for 7 days, and the expression of α-SMA protein was weak. Most of the lung fibroblasts (P-Fib) isolated from the lungs of mice injected with PLM expressed α-SMA after 7 days of culture. In the presence of PLM and transforming growth factor-β (TGF-β), the expression of α-SMA in P-Fib increased significantly, but basic fibroblast growth factor (bFGF) or platelet-derived growth factor (PDGF) had no such effect. Although bFGF had the strongest promoting effect on cell proliferation, while PLM and TGF-β had weaker effects, α-SMA positive cells expressed vimentin, while desmin expression was weak. In addition, the amount of TGF-β and bFGF produced by P-Fib was higher than that of N-Fib. These results indicate that PLM induces pulmonary fibrosis by inducing fibroblast differentiation into α-SMA positive MF, while bFGF and TGF-β play key roles in different stages of PLM-induced pulmonary fibrosis by inducing fibroblast proliferation and transformation, respectively. [4]

C61H88N18O21S2

1473.59

1472.581

C, 49.72; H, 6.02; N, 17.11; O, 22.80; S, 4.35

68247-85-8

70384-29-1 (sulfate); 68247-85-8;

6852373

Typically exists as solid at room temperature

1.6±0.1 g/cm3

1.689

-6.7

21

32

38

102

2740

20

NC([C@H](CN[C@H](C1N=C(N)C(C)=C(C(N[C@@H]([C@@H](O[C@@H]2O[C@@H](CO)[C@@H](O)[C@H](O)[C@@H]2O[C@H]2O[C@H](CO)[C@@H](O)[C@H](OC(=O)N)[C@@H]2O)C2=CN=CN2)C(N[C@@H]([C@H]([C@@H](C(N[C@H](C(NCCC2SC=C(C3SC=C(C(NCCCN[C@H](C4=CC=CC=C4)C)=O)N=3)N=2)=O)[C@H](O)C)=O)C)O)C)=O)=O)N=1)CC(=O)N)N)=O

QIMGFXOHTOXMQP-GFAGFCTOSA-N

InChI=1S/C61H88N18O21S2/c1-24-39(76-52(79-50(24)64)31(16-37(63)83)71-17-30(62)51(65)89)56(93)78-41(47(32-18-67-23-72-32)98-60-49(45(87)43(85)35(19-80)97-60)99-59-46(88)48(100-61(66)95)44(86)36(20-81)96-59)57(94)73-27(4)42(84)25(2)53(90)77-40(28(5)82)55(92)70-15-12-38-74-34(22-101-38)58-75-33(21-102-58)54(91)69-14-9-13-68-26(3)29-10-7-6-8-11-29/h6-8,10-11,18,21-23,25-28,30-31,35-36,40-49,59-60,68,71,80-82,84-88H,9,12-17,19-20,62H2,1-5H3,(H2,63,83)(H2,65,89)(H2,66,95)(H,67,72)(H,69,91)(H,70,92)(H,73,94)(H,77,90)(H,78,93)(H2,64,76,79)/t25-,26-,27+,28+,30-,31-,35-,36+,40-,41-,42-,43+,44+,45-,46-,47-,48-,49-,59+,60-/m0/s1

[(2R,3S,4S,5R,6R)-2-[(2R,3S,4S,5S,6S)-2-[(1R,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)-3-hydroxy-5-[[(2S,3R)-3-hydroxy-1-oxo-1-[2-[4-[4-[3-[[(1S)-1-phenylethyl]amino]propylcarbamoyl]-1,3-thiazol-2-yl]-1,3-thiazol-2-yl]ethylamino]butan-2-yl]amino]-4-methyl-5-oxopentan-2-yl]amino]-1-(1H-imidazol-5-yl)-3-oxopropoxy]-4,5-dihydroxy-6-(hydroxymethyl)oxan-3-yl]oxy-3,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl] carbamate

PEPLOMYCIN; Pepleomycin; 68247-85-8; Peplomycine; Peplomycin [INN]; Peplomycina; Peplomycinum; peplomicina;

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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.)