CN115531398A - Application of paclitaxel and inhibitor in antitumor drugs - Google Patents
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- CN115531398A CN115531398A CN202211224821.4A CN202211224821A CN115531398A CN 115531398 A CN115531398 A CN 115531398A CN 202211224821 A CN202211224821 A CN 202211224821A CN 115531398 A CN115531398 A CN 115531398A
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Abstract
The invention discloses an application of paclitaxel and inhibitor in antitumor drugs, and the technical scheme discloses that a paclitaxel drug composition comprises paclitaxel and minocycline combined drugs and also comprises paclitaxel and donepezil combined drugs, and the application comprises the inhibition of tumor growth and the reduction of tumor volume.
Description
Technical Field
The invention relates to the field of preparation of anti-cancer drugs, in particular to application of paclitaxel and an inhibitor in an anti-cancer drug.
Background
Paclitaxel is an antitumor drug acting on microtubules, and is mainly suitable for ovarian cancer, breast cancer and lung cancer, and has certain therapeutic effects on head and neck cancer, lymph cancer, gastric cancer, carcinoma of large intestine, melanoma, etc. From the perspective of epidemiology, lung cancer, breast cancer and gastric cancer are all high in occurrence, and the patient base numbers of the three cancers are large, so that the promotion of clinical experiments of medicaments and the research and development of medicaments are promoted.
Paclitaxel (PTX) is a diterpenoid compound extracted from bark of Taxus plant of Taxaceae, and has broad-spectrum anticancer activity and molecular formula of C 47 H 51 NO 14 The structural formula is as follows:
paclitaxel is characterized in that paclitaxel and its derivatives are widely used in breast cancer stages as main chemotherapeutic drugs, and are currently used as mainstream paclitaxel single drugs. In some reported lung cancer treatments, a combination chemotherapy regimen of paclitaxel was included in the first-line chemotherapy regimen. Therefore, the research on the pharmaceutical properties of paclitaxel is continued, and more paclitaxel drugs against other types of cancers need to be developed, and the paclitaxel combination drug is also the research and development direction of the paclitaxel drugs at present. Obviously, the inhibition effect of the single paclitaxel on the tumor cannot meet the strict treatment requirement, and a drug with better tumor inhibition effect is needed.
Disclosure of Invention
In view of the above disadvantages of the prior art, the present invention aims to provide an application of paclitaxel and an inhibitor in antitumor drugs, which has the advantages that the paclitaxel and the inhibitor are combined and synergistic, the obtained antitumor effect is superior to that of single drug administration, and the inhibition of tumor volume is significant.
The technical purpose of the invention is realized by the following technical scheme:
a paclitaxel pharmaceutical composition, comprising: comprises paclitaxel and minocycline combined drug; also includes the combination of paclitaxel and donepezil.
Further, the concentration range of the paclitaxel is 13.3-17.1 mg/kg.
Further, the concentration of the paclitaxel is 15mg/kg.
Further, the concentration range of the minocycline is 15.625-62.5 mg/kg.
Further, the concentration of the minocycline is 62.5mg/kg.
Further, the concentration range of the donepezil is 0.3 to 3mg/kg.
Further, the concentration of the donepezil is 3mg/kg.
Application of paclitaxel pharmaceutical composition in treating tumor is provided.
Further, uses include inhibiting tumor growth and reducing tumor volume.
A method for constructing a paclitaxel pharmaceutical composition comprises the following steps:
step S1: screening drugs, establishing a small-scale drug library, primarily detecting the cancer inhibition effect of different drugs combined paclitaxel under a certain concentration gradient by using an MTT (methyl thiazolyl tetrazolium) experiment, comprehensively considering an IC50 value, a drug effect, a dosage form and bioavailability, and selecting the drugs and the drug dosage to carry out the next experiment;
step S2: in vitro experiments: the drug effect can be obviously improved compared with the single use by further verifying the combination of the selected drug and the paclitaxel at the cell level, and the anti-cancer effect of the selected drug composition is proved through the judgment of indexes of cell proliferation and apoptosis.
And step S3: in vivo experiments: the mouse in vivo experiment verifies the influence of the single use of the paclitaxel and the combined use of the paclitaxel and the inhibitor on the tumor volume, the survival rate and the survival time of the mouse and the index of histological pathological scoring. .
In conclusion, the invention has the following beneficial effects:
1. paclitaxel and minocycline (minocycline, formula C) in combination 23 H 27 N 3 O 7 457.476 molecular weight and CAS number 10118-90-8), also called minocycline or mecycline, has bacteriostatic activity, belongs to PARP inhibitors, is widely applied to the treatment of acne, non-gonococcal urethritis and other infectious diseases, has pharmacological properties such as anti-inflammation, enzyme resistance, neuroprotection and the like, but has no application and clinical research report in antitumor treatment, and has synergistic effect when being used together with paclitaxelGood effect of inhibiting tumor growth and eliminating tumor.
2. Paclitaxel and donepezil in combination, donepezil (Deepezil, formula C) 24 H 29 NO 3 The molecular weight of 379.49, CAS number 120014-06-4), belongs to piperidine oxide, is a second generation specific reversible central acetylcholinesterase (AChE) inhibitor, has high selectivity to acetylcholine, is mainly used for treating mild and moderate Alzheimer Disease (AD), vascular Dementia (VD), mild cognitive dysfunction after cerebral apoplexy and other symptoms, has good tolerance, has few reports on the application of the compound in the anti-tumor field, and has good effects of inhibiting tumor growth and eliminating tumor by combining the compound with paclitaxel.
3. Animal experiments prove that the safety and the effectiveness of the combined medicament of the paclitaxel and the minocycline and the combined medicament of the paclitaxel and the donepezil in treating the tumor are high, and the potential anti-tumor potential and the clinical application value of the combined medicament of the paclitaxel and the minocycline are fully exploited.
4. The invention verifies that the donepezil and the minocycline have the medicinal value for treating the tumor, and further expands the clinical application of the donepezil and the minocycline.
Drawings
Fig. 1 is a comparative tumor plot from an in vivo experiment of the combined administration of minocycline and paclitaxel of example 2.
FIG. 2 is a comparative tumor plot of the in vivo experiment of combined donepezil and paclitaxel of example 2.
Figure 3 is a tumor contrast graph from an in vivo experiment of the combined administration of minocycline and paclitaxel of example 3.
Figure 4 is a graph of the dead tumor weight comparison of the in vivo experiment of the combined administration of minocycline and paclitaxel of example 3.
Figure 5 is a graph of body weight of nude mice from the in vivo experiment of the combined administration of minocycline and paclitaxel of example 3.
Figure 6 is a graph of tumor volume for the in vivo experiment of the combined administration of minocycline and paclitaxel of example 3.
FIG. 7 is a comparative tumor plot of the in vivo experiment of combined donepezil and paclitaxel of example 4.
Fig. 8 is a graph comparing the dead tumor weights of the in vivo experiment of the combined administration of donepezil and paclitaxel in example 4.
FIG. 9 is a graph showing the body weight of nude mice in the in vivo experiment of the combined administration of donepezil and paclitaxel in example 4.
Figure 10 is a graph of tumor volume for the in vivo experiment of donepezil and paclitaxel combination in example 4.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the embodiments of the present invention will be described in detail with reference to the accompanying drawings and the following detailed description. The advantages and features of the present invention will become more apparent from the following description.
Example 1:
a method for constructing a paclitaxel pharmaceutical composition comprises the following steps:
step S1: in the drug screening, firstly, a small-scale drug library is established by working personnel through early-stage literature search and clinical material accumulation.
Then, selecting a solvent, comparing a DMSO solvent with a solvent combination of castor oil and absolute ethyl alcohol through an MTT (methyl thiazolyl tetrazolium) experiment, finding that the castor oil and absolute ethyl alcohol have a large cytotoxicity effect on cancer cells, and neglecting the inhibition effect on the cancer cells when DMSO is maintained within 1/% o, so that DMSO is selected as the solvent for the experiment.
Then, MTT (methyl thiazolyl tetrazolium) experiments are preliminarily utilized to detect the cancer inhibition effect of paclitaxel combined with different drugs under a certain concentration gradient, the inhibition rate of 20nM paclitaxel after acting on gastric cancer cells (BG-823) for 24h, 48h and 72h is measured, the inhibition effect of 24h on the cancer cells is not obvious, the inhibition rates of 48h and 72h are obvious, but 72h is not obviously improved compared with 48h, the inhibition rate of 48h on the BG-823 gastric cancer cells is maintained at 10-20% after the paclitaxel acts on the gastric cancer cells, and the inhibition rate is improved after the paclitaxel is combined with minocycline and donepezil. The primarily selected inhibitors were minocycline and donepezil.
The structural formula of minocycline is shown as formula 2:
the structural formula of donepezil is shown in formula 3:
step S2: in vitro experiments further verify that the combination of the selected drug and the paclitaxel can obviously improve the drug effect including the influence on indexes such as cell proliferation, apoptosis and the like compared with single use at the cell level. And researches confirm that the selected medicament is dependent on which mechanism to promote the cancer inhibition effect of the paclitaxel. Only if the drug combination has its effect preliminarily judged by in vitro experiments, subsequent experiments can be carried out.
The specific experimental protocol requires:
pancreatin digests the cells in the logarithmic phase, centrifugally collects after stopping, and prepares cell suspension, the cell counting concentration is 5-10 multiplied by 104/ml, and the cell density to be detected is 5000-10000/hole.
The drug combination feasibility evaluation criterion is as follows: (1) if the Cl value can be calculated, the change rate of the inhibition rate is more than 20 percent and the Cl is less than 0.85; (2) if the Cl value is not calculated, the change rate of the inhibition rate is more than 20%. If any of the above criteria is met, it is judged that there is a synergistic effect with paclitaxel.
And step S3: in vivo experiments:
experiments in mice prove that the single-use and the combined-use of the paclitaxel have influence on indexes such as tumor volume, survival rate and survival time of the mice, and histological pathological evaluation. And verifying whether the signal pathway screened by the in vitro experiment also plays a key role in the in vivo experiment, and further evaluating the clinical potential of the research by comparing a database or detecting the expression level of the key protein of the signal pathway in the collected human samples.
The Maximum Tolerated Dose (MTD) of Paclitaxel (PTX) in mice is determined to be 15mg/kg by preliminary experiments, the number of cells inoculated in the mice in the subsequent experiments is 1-10 × 106, and the observation of the subsequent experiments shows that the cell inoculation number does not affect the growth speed of the tumor and only affects the tumor generation speed.
Example 2:
inoculating subcutaneous transplantation tumor: 6 weeks old Balb/c Nu male mice. BG-823 stomach cancer cells were diluted to 1X 10 7 Injecting 0.2ml of each nude mouse into armpit, namely inoculating 2X 10 axilla of each nude mouse respectively 6 BG-823 cells, observing the growth of transplanted tumor every other day, and making the volume of the tumor to be transplanted subcutaneously be up to 100mm 3 Administration is started.
Preparation of administration: paclitaxel, minocycline, and donepezil were diluted to dosing concentrations using normal saline.
Data collection: data collection was performed every other day, tumor volume and body weight of mice were measured, and nude mice were sacrificed 20 days after dosing and samples were collected.
The results are as follows:
(1) Minocycline and paclitaxel were administered in combination.
Dividing the nude mice grafted with the stomach cancer subcutaneous transplantation tumor into 5 groups, wherein each group comprises 3-4 nude mice (1) and normal saline; (2) paclitaxel 15mg/kg; (3) minocycline 15.625mg/kg + paclitaxel; (4) minocycline 31.25mg/kg + paclitaxel; (5) and minocycline 62.5mg/kg + paclitaxel. The administration was once every 4 days for 2 times. Nude mice were sacrificed and tumors were removed 20 days after the end of the administration, and the results are shown in fig. 1 for comparison.
And (4) conclusion: minocycline and paclitaxel have good effect of eliminating gastric cancer tumors.
(2) Donepezil and paclitaxel are co-administered.
Dividing the nude mice grafted with the stomach cancer subcutaneous transplantation tumor into 5 groups, wherein each group comprises 3-4 nude mice (1) and normal saline; (2) paclitaxel 15mg/kg; (3) donepezil 0.3mg/kg + paclitaxel; (4) 1mg/kg of donepezil and paclitaxel; (5) donepezil 3mg/kg + paclitaxel. The administration was once every 4 days for 2 times. Nude mice were sacrificed and tumors were removed 20 days after the end of the administration, and the results are shown in fig. 2 for comparison.
And (4) conclusion: donepezil and paclitaxel have good effect of eliminating gastric cancer tumor.
Example 3:
inoculating subcutaneous transplantation tumor: 6 weeks old Balb/c Nu male mice. BG-823 stomach cancer cells were diluted to 1X 10 7 Injecting 0.2ml of the vaccine into the armpit of each nude mouse per one dose, namely inoculating 2X 10 vaccine into the armpit of each nude mouse 6 BG-823 cells, observing the growth of transplanted tumor every other day, and the average volume of the tumor to be transplanted subcutaneously is 300mm 3 Administration is started.
Preparation of administration: paclitaxel, minocycline, and donepezil were diluted to dosing concentrations using normal saline.
Data collection: data collection was performed every other day, tumor volume and body weight of mice were measured, and nude mice were sacrificed 20 days after dosing and samples were collected.
(1) Minocycline and paclitaxel were administered in combination.
Dividing the nude mice grafted with the stomach cancer subcutaneous transplantation tumor into 4 groups of 3 mice, and respectively administering the nude mice (1) and normal saline to each group; (2) 62.5mg/kg minocycline; (3) paclitaxel 15mg/kg; (4) and minocycline 62.5mg/kg + paclitaxel. The administration was once every 4 days for 4 times. At the end of the administration, the nude mice were sacrificed and the tumors were removed as shown in fig. 3.
As shown in fig. 4, the placebo, paclitaxel, minocycline, and paclitaxel were co-administered and nude mice sacrificed to obtain tumor weight comparisons.
As shown in fig. 5, the first administration is taken as day 0, and the body weight of the nude mice is recorded every 4 days and plotted as fig. 5, and the body weight of the nude mice is compared with the body weight of the blank control, paclitaxel, minocycline, and minocycline taken in combination with paclitaxel.
As shown in figure 6, the first dose was taken as day 0, and tumor volumes were recorded every 4 days thereafter and plotted as figure 6, giving a comparison of tumor volumes in nude mice with placebo, paclitaxel, minocycline, and paclitaxel in combination, where p < 0.05 and p < 0.01.
And (4) conclusion: the minocycline and the paclitaxel have obvious inhibition effect on the tumor volume, and the combined use of the minocycline and the paclitaxel has the tumor inhibition effect obviously superior to that of a single medicine.
(2)
Donepezil is co-administered with paclitaxel.
Dividing the nude mice grafted with the stomach cancer subcutaneous transplantation tumor into 4 groups of 3 mice, and respectively administering the nude mice (1) and normal saline to each group; (2) donepezil 3mg/kg; (3) paclitaxel 15mg/kg; (4) donepezil 3mg/kg + paclitaxel. The administration was once every 4 days for 4 times. At the end of the administration, the nude mice were sacrificed and the tumors were removed as shown in fig. 7.
As shown in fig. 8, the blank control, paclitaxel, donepezil, and donepezil in combination with paclitaxel were sacrificed and tumor weights were compared.
As shown in fig. 9, the weight of nude mice was recorded every 4 days after the first administration was recorded as day 0, and plotted as fig. 9, to obtain the weight comparison of nude mice with blank control, paclitaxel, donepezil, and combination of donepezil and paclitaxel.
As shown in fig. 10, the first dose is recorded as day 0, and the tumor volumes are recorded every 4 days thereafter to plot as fig. 10, which is compared to the tumor volumes of nude mice given the combination of placebo, paclitaxel, donepezil, minocycline, and paclitaxel.
And (4) conclusion: the donepezil and the paclitaxel have obvious inhibition effect on the tumor volume, and the combined inhibition effect of the donepezil and the paclitaxel is obviously better than that of a single medicine.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A paclitaxel pharmaceutical composition, comprising:
comprises the combination of paclitaxel and minocycline;
also includes the combination of paclitaxel and donepezil.
2. The paclitaxel pharmaceutical composition according to claim 1, wherein: the concentration range of the paclitaxel is 13.3-17.1 mg/kg.
3. The paclitaxel pharmaceutical composition according to claim 2, wherein: the concentration of the paclitaxel is 15mg/kg.
4. The paclitaxel pharmaceutical composition according to claim 1, wherein: the concentration range of the minocycline is 15.625-62.5 mg/kg.
5. The paclitaxel pharmaceutical composition according to claim 4, wherein: the concentration of minocycline is 62.5mg/kg.
6. The paclitaxel pharmaceutical composition according to claim 1, wherein: the concentration range of the donepezil is 0.3-3 mg/kg.
7. The paclitaxel pharmaceutical composition according to claim 6, wherein: the concentration of the donepezil is 3mg/kg.
8. Use of the paclitaxel pharmaceutical composition according to any one of claims 1-7 for treating tumors.
9. Use according to claim 8, characterized in that: the uses include inhibiting tumor growth and reducing tumor volume.
10. A method of constructing a paclitaxel pharmaceutical composition according to any of claims 1-7, comprising the steps of:
step S1: screening drugs, establishing a small-scale drug library, primarily detecting the cancer inhibition effect of different drugs combined paclitaxel under a certain concentration gradient by using an MTT (methyl thiazolyl tetrazolium) experiment, comprehensively considering an IC50 value, a drug effect, a dosage form and bioavailability, and selecting the drugs and the drug dosage to carry out the next experiment;
step S2: in vitro experiments: the drug effect can be obviously improved compared with the single use of the combination of the selected drug and the paclitaxel by further verifying the cell level, and the cancer inhibition effect of the selected drug composition is confirmed by the research through judging indexes of cell proliferation and apoptosis.
And step S3: in vivo experiments: the mouse in vivo experiment verifies the influence of single-use paclitaxel and combined use of paclitaxel and inhibitor on tumor volume, survival rate and survival time of mice, and histological pathological scoring index.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105792823A (en) * | 2013-11-01 | 2016-07-20 | 皮特尼制药股份有限公司 | Pharmaceutical combinations for the treatment of cancer |
| WO2020150584A1 (en) * | 2019-01-18 | 2020-07-23 | Children's Medical Center Corporation | Compositions and methods for inducing or supplementing socs3 to abrogate tumor growth and proliferative retinopathy |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105792823A (en) * | 2013-11-01 | 2016-07-20 | 皮特尼制药股份有限公司 | Pharmaceutical combinations for the treatment of cancer |
| WO2020150584A1 (en) * | 2019-01-18 | 2020-07-23 | Children's Medical Center Corporation | Compositions and methods for inducing or supplementing socs3 to abrogate tumor growth and proliferative retinopathy |
Non-Patent Citations (2)
| Title |
|---|
| MARIE SELVY等: "Analgesic and preventive effects of donepezil in animal models of chemotherapy-induced peripheral neuropathy: Involvement of spinal muscarinic acetylcholine M2 receptors", 《BIOMEDICINE & PHARMACOTHERAPY》, vol. 149, pages 1 - 11 * |
| 曹娟: "EGCG抑制人胃腺癌SGC-7901细胞裸鼠移植瘤生长及新生血管形成实验研究", 《中国优秀博硕士学位论文全文数据库 (硕士) 医药卫生科技辑》, no. 11, pages 072 - 198 * |
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