CN103340870A - Application of andrographolide and 5-fluorouracil combined medicine in preparation of medicine for treating colon cancer - Google Patents
Application of andrographolide and 5-fluorouracil combined medicine in preparation of medicine for treating colon cancer Download PDFInfo
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- CN103340870A CN103340870A CN2013103295166A CN201310329516A CN103340870A CN 103340870 A CN103340870 A CN 103340870A CN 2013103295166 A CN2013103295166 A CN 2013103295166A CN 201310329516 A CN201310329516 A CN 201310329516A CN 103340870 A CN103340870 A CN 103340870A
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- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 title claims abstract description 33
- BOJKULTULYSRAS-OTESTREVSA-N Andrographolide Chemical compound C([C@H]1[C@]2(C)CC[C@@H](O)[C@]([C@H]2CCC1=C)(CO)C)\C=C1/[C@H](O)COC1=O BOJKULTULYSRAS-OTESTREVSA-N 0.000 title claims abstract description 29
- ASLUCFFROXVMFL-UHFFFAOYSA-N andrographolide Natural products CC1(CO)C(O)CCC2(C)C(CC=C3/C(O)OCC3=O)C(=C)CCC12 ASLUCFFROXVMFL-UHFFFAOYSA-N 0.000 title claims abstract description 28
- 229960002949 fluorouracil Drugs 0.000 title claims abstract description 18
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Abstract
The invention belongs to the field of pharmacies, and in particular relates to application of andrographolide and 5-fluorouracil combined medicine in preparation of a medicine for treating colon cancer. The application of andrographolide and 5-fluorouracil combined medicine in preparation of the medicine for treating colon cancer is disclosed by the invention for the first time; novel application of the compound is expanded.
Description
Technical field
The invention belongs to pharmaceutical field, particularly andrographolide enhanced sensitivity 5-fluorouracil (5-FU) is controlled in preparation
Treat the application in the colon cancer medicine.
Background technology
Colorectal cancer is common malignant neoplasm in digestive tract, and incidence rate is only second to gastric cancer and esophageal carcinoma.In China's common cancer death, colorectal cancer patients accounts for the 5th the male, and the women accounts for the 6th.Recent two decades comes the sickness rate of colorectal cancer increasing gradually, simultaneously, and its age of onset trend aging.At western developed country, colorectal cancer is the second malignant neoplasm that is only second to pulmonary carcinoma.Chemotherapy is the main Therapeutic Method that remains inoperable local progressive stage, recurrence or transitivity colorectal cancer at present.5-FU plays a significant role in the treatment of colon cancer as basic medicine, even but combined chemotherapy, remission rate also can only reach 40 – 50%.
Andrographolide (Andrographolide), it is the effective constituent that mainly contains of natural plants Herba Andrographis (Andrographis Paniculata (Burm.f) Ness), have the thermal detoxification of dispelling, the effect of anti-inflammatory analgetic has special efficacy to bacillary with viral upper respiratory tract infection and dysentery.Report about andrographolide and 5-FU combined effect effect does not occur as yet.The present invention has studied the effect of andrographolide in enhanced sensitivity 5-FU treatment colon cancer in vivo.
The structure of andrographolide is as follows:
Summary of the invention
Goal of the invention
Propose the application of andrographolide enhanced sensitivity 5-FU in preparation treatment colon cancer medicine, increased the indication of this chemical compound, for old medicine newly with foundation is provided.
Technical scheme
The application in preparation treatment colon cancer medicine of andrographolide and 5-fluorouracil drug combination.
Beneficial effect
The 5-FU(5-fluorouracil) as antimetabolite, at present as a line medicine for the treatment of chemotherapy of tumors.But the same with most of chemotherapeutics, have side effect such as bone marrow depression, gastrointestinal reaction, alopecia, limited its application.Present clinical many and other drug drug combination, potentiation and reduction toxic and side effects together.The present invention adopts andrographolide and 5-fluorouracil drug combination treatment colon cancer.Found through experiments with the 5-FU coupling and compare with 5FU with single, tumor suppression has improved 16.74%(58.10%vs41.36%, P＜0.05, Mann-Whitneytest) (tumor control rate=1-experimental group gross tumor volume/matched group gross tumor volume).The andrographolide list after list is compared andrographolide and 5FU coupling with 5FU, can obviously increase the area of neoplasm necrosis with not influencing cell proliferation, suppresses tumor proliferation (p-STAT3 and Ki-67 express obviously and descend).
Reduced the 5-FU consumption by drug combination in a word, thereby lowered toxic and side effects, increased the effect of resistive connection intestinal cancer, illustrated that andrographolide and 5-FU produce synergism.
Description of drawings
Fig. 1, mice-transplanted tumor change in volume.Andrographolide 5mg/kg singly uses not influence of tumor growth, but when with the 25mg/kg5FU administering drug combinations, can effectively suppress the growth of mice-transplanted tumor, andrographolide associating 5FU group is compared with 5FU with single, tumor control rate has improved 16.74%(58.10%vs41.36%, P＜0.05, Mann-Whitneytest) (tumor control rate=1-experimental group gross tumor volume/matched group gross tumor volume).
Fig. 2, mice body weight change.
Andrographolide does not have obvious influence to the mice body weight.
The mouse transplanting tumor that Fig. 3, administration divest after stopping.
Fig. 4, tumor weight.
The weighing tumor weight, the coupling group is respectively (2.518 ± 0.274) vs(3.686 ± 0.365 with single with 5FU group average tumor weight) g(P＜0.05, Students ' ttest).
Fig. 5, tumor tissues HE dyeing and immunohistochemical staining.
HE dyes after andrographolide and 5FU share as can be seen, can increase the ratio of apoptotic cell.Immunohistochemistry technology has investigated index p-STAT3 and the Ki-67 of tumor proliferation, can find out, the andrographolide list is with influencing cell proliferation, but with the 5FU coupling after, can obviously strengthen the ability that 5FU suppresses tumor cell proliferation.
The specific embodiment
1, cell culture
Mouse colonic cell is CT26, is incubated at 37 ℃, 5%CO with DMEM complete medium (containing 10%FBS)
2In the incubator of saturated humidity.
2, mouse junction cancer subcutaneous transplantation model
With the mouse colonic cell CT26 trypsinization of exponential phase, the centrifugal 5min of 250g washes twice with the PBS of pre-cooling, and is diluted to 5*10
7The concentration of/ml.Age in 6-8 week, the Balb/c mice will be scraped totally together with the right side of body hair near the axillary fossa of right side, carried out subcutaneous injection with the volume of every mice 100 μ l.After treating that tumor grows, use vernier caliper measurement tumor size every three days.Gross tumor volume computing formula: V=0.5*L1* (L2)
2, L1 represents length of tumor, and L2 represents the tumor width.
3, dosage regimen
When most of mouse tumor volume reaches 100-200mm3, be divided into four groups at random by the big young pathbreaker mice of tumor, matched group, 5FU group, andrographolide group, 5FU group and andrographolide coupling group, 15 every group.5FU25mg/kg, intraperitoneal injection, weekly twice, andrographolide sulfonated bodies 5mg/kg, intraperitoneal injection, once a day.Matched group gives PBS.
4, HE dyeing
After tumor is taken off, cut about 0.5cm, liquid-solid fixed 24 hours of 10% neutral buffered formalin or bouin, decolouring dehydration, transparent, saturating wax, embedding: 70% ethanol 30min, 80% ethanol 30min, 80% ethanol 30min, 85% ethanol 30min, 90% ethanol 30min, 95% ethanol 30min, 100% ethanol 30min * 2, ethanol: dimethylbenzene (1:1) 30min; The transparent 10min of dimethylbenzene; Dimethylbenzene: paraffin (1:1) 30min, paraffin 1h(can extend to 2 hours).Section generally about 5 μ m, 37 ℃ of roasting sheet 3h.Dimethylbenzene dewaxing 5min*3 time, 95%, 70%, 30% ethanol each 2 minutes, warm water 2 minutes.Drip the haematoxylin dye liquor or slide is immersed in the haematoxylin dye liquor, dye about 1 minute, washing 30-60 second, Yihong dyeing 30 seconds, gradient ethanol and dimethylbenzene dewater (30%, 70%, 95%, dehydrated alcohol, dimethylbenzene), dry the mounting microscopy naturally.
5, immunohistochemistry technology
Immunohistochemical assay carries out according to the description of instant second filial generation SABC ElivisionTMplus wide spectrum test kit.Simply, 1) dewaxing and the aquation of paraffin: dimethylbenzene soaks three each 10min of section, washes per step 5min with 100%, 75% and 50% ethanol gradient, washes section 3min with water.2) citrate buffer of the reparation of antigen: 10mmol/L is cut into slices behind the ebuillition of heated in pressure cooker and is put into, and builds.Be heated to jet timing 1-2min, leave thermal source, cooling is removed out the section distilled water flushing 2 times, washes the each 3min of secondary with PBS again.3) add 3% H2O2 room temperature 10min in section, then with the PBS time each 3min that gives a baby a bath on the third day after its birth.4) remove PBS liquid, every section adds a first antibody and spends the night for 4 ℃.5) PBS each 5min that gives a baby a bath on the third day after its birth time removes PBS liquid, and every section adds a polymer reinforcing agent (reagent A) incubated at room 20min, then with the PBS time each 3min that gives a baby a bath on the third day after its birth.6) remove PBS liquid, every section adds an enzyme and marks anti-Mus polymer (reagent B) incubated at room 30min, then with PBS each 3min that gives a baby a bath on the third day after its birth time.7) remove PBS liquid, every section adds a freshly prepared DAB solution, and microscopically is observed 3-10min.8) tap water flushing, haematoxylin is redyed, 0.1% acidic alcohol differentiation, the tap water flushing, PBS returns indigo plant.Take pictures.9) cut into slices through the gradient alcohol dehydration drying neutral gum sealing.
6, statistics
Data are represented with mean ± SD.Mann-Whitneytest is used in comparison between two groups of gross tumor volumes, relatively uses Student ' stwo-tailedt-test to check difference between each group between two groups of other data.* * represents P＜0.001, and * * represents P＜0.01, and * represents P＜0.05.
Interpretation
From Fig. 1,3,4 as can be seen, the andrographolide list is used mouse junction cancer unrestraint effect, 5FU25mg/kg can significantly suppress growth of tumor, but after having united andrographolide, the inhibitory action of the tumor of 5FU obviously strengthens, when the treatment terminal point, drug combination group tumor size is compared with the 5FU group, tumor control rate has improved 16.74%(58.10%vs41.36%, P＜0.05, Mann-Whitneytest) (tumor control rate=1-experimental group gross tumor volume/matched group gross tumor volume), average tumor weight is respectively (2.518 ± 0.274) vs(3.686 ± 0.365) g(P＜0.05, Students ' ttest), the 1.168g that descended.
And as can be seen from Figure 2, andrographolide does not have obvious influence to body weight.
In addition, as can be seen from Figure 5, compare with 5FU with single, after andrographolide and the 5FU coupling, can obviously increase the area of neoplasm necrosis, suppress tumor proliferation (p-STAT3 and Ki-67 express obviously and descend).
Claims (1)
1. andrographolide and the 5-fluorouracil drug combination application in preparation treatment colon cancer medicine.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013103295166A CN103340870A (en) | 2013-07-31 | 2013-07-31 | Application of andrographolide and 5-fluorouracil combined medicine in preparation of medicine for treating colon cancer |
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| CN2013103295166A CN103340870A (en) | 2013-07-31 | 2013-07-31 | Application of andrographolide and 5-fluorouracil combined medicine in preparation of medicine for treating colon cancer |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107320486A (en) * | 2016-04-28 | 2017-11-07 | 南京大学 | Application of the andrographolide with cisplatin combined medication in treatment cisplatin-resistant lung-cancer medicament is prepared |
| CN112480097A (en) * | 2020-11-26 | 2021-03-12 | 汕头大学医学院 | Inhibitor of targeting ETS structural domain protein and application thereof |
-
2013
- 2013-07-31 CN CN2013103295166A patent/CN103340870A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 杨琼等: "穿心莲内酯联合5-FU对人胃癌BGC-823细胞的体内外抑制作用", 《中华中医药学刊》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107320486A (en) * | 2016-04-28 | 2017-11-07 | 南京大学 | Application of the andrographolide with cisplatin combined medication in treatment cisplatin-resistant lung-cancer medicament is prepared |
| CN112480097A (en) * | 2020-11-26 | 2021-03-12 | 汕头大学医学院 | Inhibitor of targeting ETS structural domain protein and application thereof |
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Application publication date: 20131009 |