CN111184868A - Application of zotara in preparation of medicine for enhancing anti-cancer activity of daunorubicin in vivo and in vitro - Google Patents
Application of zotara in preparation of medicine for enhancing anti-cancer activity of daunorubicin in vivo and in vitro Download PDFInfo
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- CN111184868A CN111184868A CN202010206074.6A CN202010206074A CN111184868A CN 111184868 A CN111184868 A CN 111184868A CN 202010206074 A CN202010206074 A CN 202010206074A CN 111184868 A CN111184868 A CN 111184868A
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- 229960000975 daunorubicin Drugs 0.000 title claims abstract description 73
- STQGQHZAVUOBTE-VGBVRHCVSA-N daunorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(C)=O)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 STQGQHZAVUOBTE-VGBVRHCVSA-N 0.000 title claims abstract description 73
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- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 3
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- 229930195725 Mannitol Natural products 0.000 claims description 3
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 3
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- ZAFYATHCZYHLPB-UHFFFAOYSA-N zolpidem Chemical compound N1=C2C=CC(C)=CN2C(CC(=O)N(C)C)=C1C1=CC=C(C)C=C1 ZAFYATHCZYHLPB-UHFFFAOYSA-N 0.000 description 2
- 229960001475 zolpidem Drugs 0.000 description 2
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Images
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/02—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/28—Mercury; Compounds thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
The invention relates to an application of zotara in preparing a medicine for enhancing the in-vivo and in-vitro anticancer activity of daunorubicin, belonging to the technical field of medicine preparation. The invention provides an application of zotara in preparing a medicine for enhancing the in vivo and in vitro anticancer activity of daunorubicin. The invention combines the daunorubicin anticancer monomer drug with the zotara nano-particles for in vitro administration, and proves that the zotara can promote the enrichment of daunorubicin in different cancer cells. Meanwhile, the apoptosis promoting effect of the medicine can be enhanced.
Description
Technical Field
The invention relates to the technical field of medicine preparation, in particular to application of zotara in preparing a medicine for enhancing the in-vivo and in-vitro anticancer activity of daunorubicin.
Background
Daunorubicin belongs to anthracycline antineoplastic drugs, and is a chemotherapeutic drug widely applied to blood system tumors and various solid tumors at present. However, daunorubicin has a large toxic and side effect while killing tumor cells. Moreover, as chemotherapy progresses, many tumor cells can resist chemotherapeutic drugs to reduce the curative effect, and multidrug resistance becomes a main reason for the ultimate failure of chemotherapy of tumor patients. Increasing the dosage of the medicine brings more toxic and side effects to patients, so that finding ways, adjuvant agents or methods which can effectively improve the concentration of the medicine at tumor parts is one of the hot spots of research in the field of tumor treatment. How to efficiently deliver the daunorubicin to tumor tissues, overcoming multidrug resistance and reducing toxic and side effects is the key to fully exert the drug effect.
Disclosure of Invention
The invention aims to provide the application of zotara in preparing a medicine for enhancing the anticancer activity of daunorubicin in vitro and in vivo. Zotara can promote the enrichment of daunorubicin in different cancer cells. Meanwhile, the apoptosis promoting effect of the medicine can be enhanced.
The invention provides an application of zotara in preparing a medicine for enhancing the in vivo and in vitro anticancer activity of daunorubicin.
The invention also provides application of zotara in preparation of a medicine for promoting daunorubicin to be enriched in cancer cells.
The invention also provides application of zotara in preparing a medicine for enhancing daunorubicin and promoting cancer cell apoptosis.
Preferably, in the application, the mass ratio of zotaro to daunorubicin is (1-500): (100 to 2000).
Preferably, the dosage forms of the medicine comprise injection and powder injection.
Preferably, the medicament further comprises pharmaceutically acceptable auxiliary materials: the injection comprises sodium hydroxymethyl cellulose; the powder for injection comprises glucose, lactose and/or mannitol.
The invention provides an application of zotara in preparing a medicine for enhancing the in vivo and in vitro anticancer activity of daunorubicin. The daunorubicin anticancer monomer drug and the zotara nano-particles are administrated in vitro, and the influence of the monomer drug and the zotara on the intracellular drug concentration change and the apoptosis promotion of HN4, K562 and HpG-2 cells is examined. It was confirmed that zotara can promote the enrichment of daunorubicin in various cancer cells. Meanwhile, the apoptosis promoting effect of the medicine can be enhanced. Test results show that the administration of the zotara and the daunorubicin can obviously increase the daunorubicin entering the interiors of 3 different cancer cells, and the intracellular drug concentration of HN4, K562 and HpG-2 cells in 24h, 48h and 72h is obviously increased; the administration of the zotara and the daunorubicin can obviously promote the apoptosis rate of HN4 cancer cells at different time periods, and the zotara has obvious synergistic effect on the daunorubicin.
Drawings
FIG. 1 is a graph showing the effect of various time periods following administration of the zotaro combination daunorubicin provided by the present invention on the enhancement of drug entry into HN4 cells;
FIG. 2 is a graph showing the effect of various times of administration of the zotaro combination daunorubicin provided by the present invention on the promotion of drug entry into the interior of K562 cells;
FIG. 3 is a graph showing the effect of various times after administration of the zotaro combination daunorubicin provided by the present invention on the promotion of drug entry into HpG-2 cells;
FIG. 4 shows that the administration of zolpidem in combination with daunorubicin for 48h increases the apoptotic effect of K562 cells;
FIG. 5 shows that administration of zolpidem in combination with daunorubicin for 72h provided by the present invention increases the apoptotic effect of K562 cells.
Detailed Description
The invention provides an application of zotara in preparing a medicine for enhancing the in vivo and in vitro anticancer activity of daunorubicin. The source of the zotara is not particularly limited in the present invention, and conventional commercial zotara sources well known to those skilled in the art can be used. Zotara can promote the enrichment of daunorubicin in different cancer cells. Meanwhile, the apoptosis promoting effect of the medicine can be enhanced, and the in-vivo and in-vitro anticancer activity of the daunorubicin can be further enhanced.
The invention also provides application of zotara in preparation of a medicine for promoting daunorubicin to be enriched in cancer cells. The administration of zotara combined with daunorubicin can remarkably increase the entry of daunorubicin into the interior of 3 different cancer cells (HN4, K562 and HpG-2), and the intracellular drug concentration of three cells of HN4, K562 and HpG-2 is remarkably increased at 24h, 48h and 72h after the administration.
The invention also provides application of zotara in preparing a medicine for enhancing daunorubicin and promoting cancer cell apoptosis. The administration of the zotara and the daunorubicin can obviously promote the apoptosis rate of HN4 cancer cells at different time periods, and the zotara has obvious synergistic effect on the daunorubicin.
In the application, the mass ratio of zotara to daunorubicin is (1-500): (100-2000), more preferably (1-500): (1000 to 2000). In the present invention, when the drug is co-administered with daunorubicin, the drug is preferably administered simultaneously with daunorubicin, or daunorubicin is administered 24 hours after the drug is administered. In the invention, when in vitro administration, the concentration of zotaro in the medicine is preferably 1-500 ng/ml, and the concentration of daunorubicin is 0.1-2.0 mu g/ml; when the medicine is administrated in vivo, the administration amount of the zotaro in the medicine is preferably 10ng/kg body weight to 200 mug/kg body weight, the zotaro is preferably administrated once every 2-7 days, the zotaro can be administrated by intraperitoneal injection, intragastric administration or intravenous administration, the administration amount of the daunorubicin is preferably 1-5 mg/kg body weight, and the daunorubicin is administrated according to a clinical administration method.
In the invention, the dosage form of the medicine preferably comprises injection and powder injection. The preparation method of the injection and the powder injection is not particularly limited, and the conventional preparation method of the injection and the powder injection, which is well known by the technical personnel in the field, is adopted.
In the invention, the medicine also comprises pharmaceutically acceptable auxiliary materials: the injection comprises sodium hydroxymethyl cellulose; the powder for injection comprises glucose, lactose and/or mannitol.
The use of zotai in the present invention for preparing a medicament for enhancing anti-cancer activity of daunorubicin in vitro and in vivo is further described in detail with reference to the following specific examples, and the technical scheme of the present invention includes but is not limited to the following examples.
Example 1
1. Zote promotes entry of daunorubicin into cells
1.1 methods of administration
Grouping and administration: the K562 leukemia cells, the HN4 oral squamous carcinoma epithelial cells and the HpG2 liver cancer cells are respectively divided into a daunorubicin single administration group of 24h, a daunorubicin single administration group of 48h and a daunorubicin combined administration group of 24h, a daunorubicin combined administration group of 48h and a daunorubicin combined administration group of 72 h.
1.2 sample collection: collecting K562 leukemia cells, HN4 oral squamous carcinoma epithelial cells and HpG2 hepatocarcinoma cells in logarithmic growth phase, and adjusting cell density to 2 × 106Respectively inoculating in culture dish, and placing in 5% CO2And culturing in an incubator at 37 ℃ until the cells grow to 70-80%. The three cells were treated with daunorubicin at a concentration of 0.25. mu.g/ml and 5ng/ml of zotara in combination with daunorubicin, washed 3 times with PBS after 24h, 48h, and 72h, and harvested separately.
1.3 sample treatment
And (3) centrifuging the cells after the daunorubicin medicament treatment and the zotara combined daunorubicin treatment for 24h, 48h and 72h for 5min by using a low-temperature centrifuge at 1000r/min, removing supernatant by suction, obtaining the collected cells as the residual precipitate, adding 50uL of cell lysate, adding 1ml of methanol for dissolving after full lysis, and detecting by HPLC.
1.4 sample analysis
Chromatographic conditions are as follows: the mobile phase is 20% methanol at equal temperature for 30 min; the flow rate is 1 mL/min; the column temperature is 30 ℃; the detection wavelength is 210 nm; the amount of sample was 10. mu.L.
2. Research for promoting synergism of daunorubicin
2.1MTT assay
The log phase K562 leukemia cells were digested with 0.25% trypsin at 5X 10 per well3The cells were seeded in 96-well culture plates and incubated at 37 ℃ with 5% CO2Cultured in an incubator. Culturing for 24h, discarding supernatant, setting PBS hole as blank group, non-drug-adding hole as control group, and adding daunorubicin with different concentrations into each hole of experimental group (treating for 48 hr)The administration concentration is 1.25 mug/ml, 2.5 mug/ml and 5 mug/ml; treatment for 72 hours dosing concentrations of 0.5. mu.g/ml, 1. mu.g/ml, 1.5. mu.g/ml) or in combination with 1ng/ml zotare treatment, 6 parallel wells per group were set. After 48 and 72h incubation, respectively, the supernatant was aspirated off, 20. mu.L of 5mg/m L MTT solution was added to each well, and CO was added2After further culturing for 4h in the incubator, the supernatant was aspirated off, 150. mu.L of dimethyl sulfoxide (DMSO) was added to each well, the mixture was slowly shaken for 10min on a 300rmp shaker at a constant temperature of 37.5 ℃ and absorbance values at 490nm and 570nm were measured using a microplate reader, and the absorbance values were adjusted to zero using blank wells. According to a cell viability calculation formula, cell viability (%) is (OD value of an experimental group/OD value of a control group) multiplied by 100%, and the average cell viability of each experimental group is calculated by taking an average value.
Administration of zotara in combination with daunorubicin can significantly increase daunorubicin entry into the interior of 3 different cancer cells. The intracellular drug concentration of HN4, K562 and HpG-2 was significantly increased at 24, 48 and 72h of administration. When the Zote and the daunorubicin are administrated in HN4 cells, the drugs are inhibited from entering the cells for 24h and 48h, but are obviously promoted to enter the cells for 72h, and the increment is 120.3 +/-12.0%. In K562 cells, the increment of the combination group at 48h and 72h was 90.4. + -. 40.7% and 32.8. + -. 27.2%, respectively, compared with that of the single administration group. The increments of 24h, 48h and 72h of zotaro in HpG2 cells to facilitate drug entry into the cells were 12.6 + -7.9%, 82.4 + -22.0% and 24.8 + -8.0%, respectively (Mean + -SD). As shown in fig. 1, 2 and 3, fig. 1 shows the promoting effect of the zotarabicin and daunorubicin on the medicine entering into the cell of HN4 at different times, fig. 2 shows the promoting effect of the zotarabicin and daunorubicin on the medicine entering into the cell of K562 at different times, and fig. 3 shows the promoting effect of the zotarabicin and daunorubicin on the medicine entering into the cell of HpG-2 at different times.
The administration of the zotara and the daunorubicin can obviously promote the apoptosis rate of HN4 cancer cells at different time periods, and the zotara has obvious synergistic effect on the daunorubicin. The administration lasts for 48h, the combined administration group of the zotaro and the daunorubicin at medium dose has obviously better apoptosis-promoting effect on K562 cells than that of the single administration group, and the increment is 33.8 +/-24.4%. The intermediate dose of zotaro had an increased pro-apoptotic effect on daunorubicin of 22.4 ± 24.8% at 72h of administration. As shown in fig. 4 and 5, respectively, fig. 4 shows that the administration of zoledram in combination with daunorubicin for 48h increases the apoptosis-promoting effect of the drug K562 cells, and fig. 5 shows that the administration of zoledram in combination with daunorubicin for 72h increases the apoptosis-promoting effect of the drug K562 cells.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (6)
1. Use of zotara in the preparation of a medicament for enhancing the anticancer activity of daunorubicin in vitro and in vivo.
2. Use of zotara in the preparation of a medicament for promoting the enrichment of daunorubicin in cancer cells.
3. Use of zotara in preparing medicine for enhancing daunorubicin and promoting cancer cell apoptosis is provided.
4. The use according to any one of claims 1 to 3, wherein the use comprises the following steps in a mass ratio of zotaro to daunorubicin (1 to 500): (100 to 2000).
5. The use of any one of claims 1 to 4, wherein the dosage form of the medicament comprises an injection and a powder injection.
6. The use of claim 5, wherein the medicament further comprises pharmaceutically acceptable excipients: the injection comprises sodium hydroxymethyl cellulose; the powder for injection comprises glucose, lactose and/or mannitol.
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Application publication date: 20200522 |