Peptides and pharmaceutical compositions comprising said peptides and medical composition for treating cancer
A composition and drug technology, applied in the direction of drug combination, medical preparations containing active ingredients, gene therapy, etc., can solve the problems that there is no gene into cancer cells, it is difficult to achieve the expected goal, and it is difficult to enhance the immune response
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Embodiment 1
[0090] Different cancer cell lines were cultured under hypoxic conditions, and the expression of adrenomedullin mRNA was detected by Northern blotting. Using a hypoxic incubator (Wakenyaku Industry), cancer cell lines were incubated in 1% O under hypoxic conditions. 2 concentration for 12 hours. Also at 20% O 2 Concentrations were cultured as a control. After culturing, RNA was extracted from different cancer cell lines with TRIZOL reagent (LIFETECHNOLOGIES). RNA (20 μg) was electrophoresed on a formaldehyde-agarose gel and subsequently hybridized with an adrenomedullin-specific probe. see results Figure 1 .
[0091] The cancer cell lines used here are listed below
[0092] KATO III gastric cancer cell line
[0093] HCT116 rectal cancer cell line
[0094] DLD1 rectal cancer cell line
[0095] KM-12 rectal cancer cell line
[0096] PC-6 Lung Cancer Cell Line
[0097] TAOV Ovarian Cancer Cell Line
[0098] PCI-10 pancreatic cancer cell line
[0099] HepG2 liver can...
Embodiment 2
[0108] Implanted subcutaneously in the dorsal side of CB17lcr-scid J1 mice (CLEA JAPAN Inc.) 10 7 PCI-43 cells. Proliferation of PCI-43 cells has been demonstrated to form tumors that exceed 5 mm in diameter after 7 days. After this confirmation, 50 µg each of adrenomedullin and the peptide of the present invention (dissolved in 0.1 ml of physiological saline) were injected into the tumor once a day from day 7 until day 16, respectively. A peptide consisting of SEQ ID NO: 2 was used as the peptide of the present invention (hereinafter also referred to as "adrenomedullin antagonist"). This peptide was purchased from Wako Pure Chemical Industries, Ltd.
[0109] Tumor diameters were observed visually every three days. see results figure 2 , which shows the tumor size for each administration of adrenomedullin, adrenomedullin antagonist and saline (V3). The horizontal axis represents the days after implantation of PCI-43 cells, and the vertical axis represents the tumor volume...
Embodiment 3
[0112] Implanted subcutaneously in the dorsal side of CB17lcr-scid J1 mice (CLEA JAPAN Inc.) 10 7 PCI-43 cells. Proliferation of PCI-43 cells has been demonstrated to form tumors that exceed 5 mm in diameter after 7 days. After this confirmation, 50 μg each of adrenomedullin and adrenomedullin antagonist (dissolved in 0.1 ml of normal saline) were injected into the tumor every three days from the seventh day, i.e., on day 7, Day 10, Day 13 and Day 16. The adrenomedullin and adrenomedullin antagonist used here were the same as those used in Example 1.
[0113] On the 21st day, the mice were sacrificed, and the tumors were excised and weighed. Photos of the excised tumor are shown in image 3 , tumor weighing results see Figure 4 .
[0114] From image 3 It was clearly seen that in the adrenomedullin antagonist-administered group, the tumor size became smaller than that in the adrenomedullin-administered group. In addition, if Figure 4 As clearly shown, the tumor weig...
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