CN116603063A - New use of proton pump inhibitor - Google Patents

New use of proton pump inhibitor Download PDF

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Publication number
CN116603063A
CN116603063A CN202310419576.0A CN202310419576A CN116603063A CN 116603063 A CN116603063 A CN 116603063A CN 202310419576 A CN202310419576 A CN 202310419576A CN 116603063 A CN116603063 A CN 116603063A
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proton pump
pump inhibitor
rabeprazole
multiple myeloma
cells
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刘丽宏
宫丽丽
成虎
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China Japan Friendship Hospital
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • 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
    • A61K31/69—Boron compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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Abstract

The invention discloses a new application of a proton pump inhibitor, namely an application of the proton pump inhibitor in preparing a medicament for treating multiple myeloma. Experiments prove that the rabeprazole, the omeprazole, the pantoprazole, the ilaprazole, the lansoprazole, the dexlansoprazole, the tenatoprazole, the leminoprazole, the Sha Weila oxazole, the picoprazole, the temozolomide and the esomeprazole can inhibit the activity of multiple myeloma cells at a low concentration on a cellular level, the rabeprazole can inhibit the activity of the multiple myeloma cells at an animal level at a dosage not exceeding clinical dosage, and the proton pump inhibitors can inhibit the multiple myeloma cells at the cellular level and the animal level based on the proton pump inhibitors, so that the rabeprazole can be used for preparing medicines for treating the multiple myeloma.

Description

New use of proton pump inhibitor
Technical Field
The invention relates to the field of medicine. More particularly, the present invention relates to a novel use of a proton pump inhibitor.
Background
Multiple myeloma is the second most common hematological malignancy. It is a plasma cell malignancy, most of which originate from unknown monoclonal immunoglobulin blood, develop into smoky multiple myeloma, and finally develop into newly diagnosed multiple myeloma. Multiple myeloma can bring low erythrocyte anemia, hypercalcemia, fracture and other complications to patients, and abnormal accumulation of M protein can cause kidney damage to the patients, and immune disorder can also increase infection risk of the patients, so that the damage to the patients is great.
In recent decades, with the advent of autologous stem cell transplantation, the use of new drugs such as immunomodulatory drugs and proteasome inhibitors, the improvement of multiple myeloma treatment changed the natural course of the disease and prolonged the survival of the patient. But eventually patients experience multiple relapses, with shorter and shorter remissions, ultimately dying from multiple myeloma or treatment-related complications. The prognosis of multiple myeloma survival remains still optimistic, and it is an urgent task to try new drugs. Therefore, the invention in the field has great social and economic significance.
Proton pump inhibitors are currently used primarily in the treatment of diseases requiring reduced gastric acid secretion, such as erosive or ulcerative gastroesophageal reflux disease, non-erosive reflux disease, duodenal and gastric ulcers, and pathological hypersecretion diseases, including Zollinger-Ehrlich syndrome, and in addition to eradication of helicobacter pylori. Proton pump inhibitors have recognized therapeutic effects and safety in the treatment of gastric acid related diseases.
Disclosure of Invention
The invention aims to provide a new application of a proton pump inhibitor in preparing a medicament for treating multiple myeloma.
To achieve these objects and other advantages and in accordance with the purpose of the invention, a novel use of a proton pump inhibitor for preparing a medicament for treating multiple myeloma is provided.
Preferably, the proton pump inhibitor comprises rabeprazole, omeprazole, pantoprazole, ilaprazole, lansoprazole, dexlansoprazole, tenatoprazole, leminoprazole, sha Weila oxazole, picoprazole, temozolomide and esomeprazole, and a compound preparation containing the effective components of the corresponding proton pump inhibitor.
Preferably, the proton pump inhibitor is rabeprazole, and the use of the rabeprazole and bortezomib in combination for preparing medicines for treating multiple myeloma.
Preferably, the medicament for treating the multiple myeloma further comprises a pharmaceutically acceptable carrier or a pharmaceutical auxiliary material of the proton pump inhibitor.
Preferably, the dosage forms of the medicament for treating the multiple myeloma comprise oral preparations and injection preparations.
Preferably, the oral formulation comprises: tablets, capsules, oral liquids, buccal tablets, granules, suspension, dripping pills, sustained release agents, controlled release agents and targeting preparations.
Preferably, the injection preparation comprises: injection and powder injection.
The invention at least comprises the following beneficial effects: experiments prove that the rabeprazole, the omeprazole, the pantoprazole, the ilaprazole, the lansoprazole, the dexlansoprazole, the tenatoprazole, the leminoprazole, the Sha Weila oxazole, the picoprazole, the temozolomide and the esomeprazole can inhibit the activity of multiple myeloma cells at a low concentration on a cellular level, the rabeprazole can inhibit the activity of the multiple myeloma cells at an animal level at a dosage not exceeding a clinical dosage, and the proton pump inhibitors can inhibit the multiple myeloma cells at the cellular level and the animal level based on the proton pump inhibitors, so that the rabeprazole can be used for preparing medicaments for treating the multiple myeloma.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.
Drawings
FIG. 1 is a schematic diagram showing the combination of UAP1 and rabeprazole in example 1 of the present invention;
FIG. 2 is a graph showing the experimental results of UAP1 expression level and rabeprazole efficacy in example 2 of the present invention;
FIG. 3 is a graph showing the experimental results of experiments for measuring cell viability by CCK8 assay performed in example 3 of the present invention;
FIG. 4 is a graph showing the experimental results of the CCK8 assay cell viability measurement experiment performed in example 4 of the present invention;
FIG. 5 is a graph showing the experimental results of the CCK8 assay cell viability measurement experiment performed in example 5 of the present invention;
FIG. 6 is a graph showing the experimental results of experiments for measuring cell viability by CCK8 assay performed in example 6 of the present invention;
FIG. 7 is a graph of experimental results of subcutaneous xenograft experiments performed in accordance with example 7 of the present invention (each experimental group contains 3 samples);
FIG. 8 is a graph of experimental results of subcutaneous xenograft experiments performed in example 8 of the present invention (each experimental group contains 3 samples);
FIG. 9 is a graph showing the experimental results of experiments for measuring cell viability by CCK8 assay performed in example 9 of the present invention;
FIG. 10 is a graph showing the experimental results of the CCK8 assay cell viability measurement experiment performed in example 10 of the present invention;
FIG. 11 is a graph showing the experimental results of experiments for measuring cell viability by CCK8 assay conducted in example 11 of the present invention;
FIG. 12 is a graph showing the experimental results of the CCK8 assay cell viability measurement experiment performed in example 12 of the present invention;
FIG. 13 is a graph showing the experimental results of experiments for measuring cell viability by CCK8 assay conducted in example 13 of the present invention;
in fig. 2 to 6 and 9 to 13, mean±sem, P < 0.001, and P < 0.0001.
Detailed Description
The present invention is described in further detail below with reference to the drawings to enable those skilled in the art to practice the invention by referring to the description.
The experimental methods described in the following embodiments are conventional methods unless otherwise indicated, and the reagents and materials are commercially available.
The main reagents and materials in the following embodiments include: rabeprazole, omeprazole, pantoprazole, ilaprazole, lansoprazole, dexlansoprazole, tenatoprazole, leminoprazole, sha Weila oxazole, picoprazole, temozolomide and esomeprazole (purchased from MCE), PBS, fetal bovine serum, medium (purchased from Gibco, usa), CCK8 reagent (purchased from beijing biotech), NCI-H929 cells (purchased from the pool of synergetic cells);
animal origin in the following embodiments: beijing Vitolihua laboratory animal technology Co., ltd;
animal breed strain: mice, NOG; gender: a male; week-old: 5 weeks of age; weight of: 18-22 g.
Animals need to be adapted to the environment for 1-7 days before the test, healthy animals are selected as the tested animals, random grouping is carried out according to the weight, and after grouping, each animal is assigned a unique and permanent animal number. IVC cages were bred at a density of 5 animals per cage, with the litter replaced weekly.
Experimental animal house requirements in the following embodiments: room temperature is 20-26 ℃; the relative humidity is 40-70%; automatic illumination, 12h light and shade alternation (07 points 00 separate lamps, 19 points 00 separate lamps off); the standard of the animal room meets the national standard GB 14125-2010 of the people's republic of China; and (3) drinking water: RO membrane reverse osmosis sterile water.
The test method in the following embodiments comprises:
A. molecular docking drug screening based on UAP1
And downloading a protein structure file encoded by the gene by using a PDB database, and downloading an FDA approved small molecule drug structure by using a ZINC database and a drug Bank database. Virtual screening was performed by docking all prepared ligands at defined active sites using LibDock module in Discovery studio software.
B. Sample collection and determination of UAP1 expression level
1) Cell preparation: taking all groups of cells in logarithmic growth phase, blowing the cells into single cells by a rubber head dropper, and suspending the cells in a cell growth medium of 10% fetal calf serum for later use;
2) Inoculating cells: NCI-H929 cells were seeded in six well plates, each set with three complex wells;
3) Cell dosing: the experimental groups are respectively added with rabeprazole with different concentrations, and the control holes are added with PBS with the same volume;
4) After 48h, cell samples were collected, RNA was extracted and UAP1 expression was determined.
C. cDNA Synthesis
1) Taking out the RNA extracted in the step B from a refrigerator at the temperature of-80 ℃ and placing the RNA on ice for thawing;
2)5×g DNA Buffer、FQ-RT Primer Mix、10×King RT Buffer，RNase-Free ddH 2 thawing O at room temperature (15-25deg.C);
3) Vortex shaking and mixing each reagent in 2), and briefly centrifuging.
4) According to the gDNA removal reaction system (5 XgDNA Buffer 2. Mu.L; a proper amount of Total RNA; RNase-Free ddH 2 O is added to 10 mu L) to prepare a mixed solution;
5) Incubating for 3min at 42 ℃ by a PCR instrument, and then placing on ice;
6) A reverse transcription reaction system Mixture (MIX) was prepared according to the kit instructions. Reverse transcription reaction system: 10 XKing RT Buffer 2. Mu.L; fast King RT Enzyme mix 1 μL; FQ-RT Primer mix 2. Mu.L; RNase-Free ddH 2 O is added to 10 mu L;
7) MIX in reverse transcription reaction, add MIX to gDNA removal reaction system mixture, MIX well by vortex oscillation, centrifuge briefly.
8) Incubating for 15min at 42 ℃ by a PCR instrument; incubation was performed at 95℃for 3min. The obtained cDNA can be used for subsequent experiments or stored in a low-temperature refrigerator at-80 ℃ for standby.
D. Real-time fluorescent quantitative PCR
1) After cDNA synthesis, 180. Mu.L of RNase-Free ddH was added before use 2 O; (dilution of cDNA reduces errors, but should not be diluted when applied to the lower level of gene expression
2) Fluorescent reaction system: 10. Mu.L 2X SuperReal Pre Mix Plus, 0.6. Mu.L forward primer, 0.6. Mu.L reverse primer, 2. Mu.L cDNA, 18. Mu.L ddH 2 O. After the reaction system is prepared, vortex oscillation and uniform mixing are carried out, and centrifugation is carried out briefly.
3) Real-time fluorescent quantitative PCR instrument reaction procedure: 95 ℃ for 15min;95℃10s,60℃20s,72℃32s for a total of 40 cycles.
E. CCK8 measurement cell viability assay, specific methods and procedures:
1) Cell preparation: taking all groups of cells in logarithmic growth phase, blowing the cells into single cells by a rubber head dropper, and suspending the cells in a cell growth medium of 10% fetal calf serum for later use;
2) Inoculating cells: the cells of interest were seeded at a density in 96-well plates with 100 μl of medium per well, 4 multiplex wells per experimental set. Blank wells (medium only), control wells (seed cells only) were also set.
3) Cell dosing: the experiment is respectively added with the medicaments to be detected with different concentrations, the control hole is added with the culture medium with the same volume, and the blank hole is not added with other substances;
4) Transferring the 96-well plate into an incubator for culturing, wherein the environment in the incubator is 37 ℃ and 5 percent of CO 2 ；
5) Experimental group, blank well and control well, 10. Mu.L CCK8 was added to each well and placed in incubator (37 ℃,5% CO) 2 ) Incubate for 1-4 hours.
F. Subcutaneous xenograft experiments, specific methods and procedures:
1) Animal preparation: NOG mice were adaptively bred in SPF-grade environment for one week;
2) Cell preparation: taking NCI-H929 cells in a logarithmic growth phase, centrifuging at 1000rpm for 5min, washing with PBS for 2 times, adding PBS to resuspend the cells, and counting for later use;
3) Animal preparation: the axilla of NOG mice on the day of inoculation are dehaired;
4) Transferring the cells to the place where the animals are located as soon as possible, ensuring that they remain on ice for viability at all times;
5) The cell suspension was gently mixed to ensure that the single cell suspension could be homogenized by pipetting up and down slowly several times using a P1000 pipette. This was repeated at inoculation to ensure a uniform cell suspension was injected each time;
6) The use of a large gauge needle (e.g., an 18 gauge needle) to aspirate cells into the syringe reduces cell damage due to shear forces during injection. Wiping the injection area with an alcohol cotton ball to perform side injection;
7) 6.5X10 mice were vaccinated with each 6 week old male NOG mouse 6 The NCI-H929 cells were subcutaneously injected, taking care to ensure that the cells were injected correctly. If the needle is too shallow, tumor cells may be injected into the skin (intradermal injection), while if too deep, tumor cells may be injected into (or pass through) the muscle layer under the skin, severely leading to failure of the neoplasia;
8) Ensuring that the cell injection (typically 0.1 mL) is smooth and discreet, not in a hurry, and completed in a few seconds;
9) After the injection is completed, the needle is slowly withdrawn, ensuring that no cell suspension is spilled at the wound. A small tumor or swelling will form at the site of tumor cell injection;
10 After the cells are injected, the mice are put into fresh cages in time, so that the mice can eat at any time
Object and water. Observing the mice for 30min after injection, and ensuring that each mouse is recovered to be normal;
11 Mice were observed daily after inoculation and dosing was started after access to the pads (approximately 96h post inoculation). The administration mode is intraperitoneal injection. Carbon dioxide was used for euthanasia at day 21 post inoculation and the material was obtained. Other innocent treatment of animal carcasses. All experiments were performed according to the Chinese animal welfare method and approved by the authorities.
Example 1: UAP 1-based drug virtual screening
And downloading the protein structure file of UAP1 by using the PDB database, and downloading the compound database from the drug Bank database. Qualitative pharmacophore models were generated using Discovery Studio for database screening to determine new scaffolds for future drug Discovery. Pharmacophore modeling is often closely related to the docking procedure, the first step being to flexibly arrange ligand molecules into a rigid macromolecular environment, and then to estimate the closeness of interactions by different scoring functions. Docking takes all information from the rigid protein environment and scores several possible interaction patterns for different alignments. The ligand binding pocket region of UAP1 is selected as a binding site to screen for compounds that may inhibit UAP1, and the "prepare ligand" module is used to prepare the screened active ingredient to obtain an effective trisection conformation. After removal of the crystal water molecules, the multi-conformation of the target protein is removed using a "protein preparation" module, supplementing the incomplete amino acid residues. Subsequently, molecular docking was performed in a "LibDock" module, and LibDock score was required to evaluate the affinity of the target protein and active ingredient.
Fig. 1 is a schematic diagram showing the binding of rabeprazole candidate to UAP 1. Rabeprazole can bind to UAP1 to exert an anti-multiple myeloma effect.
Example 2: UAP1 expression level and rabeprazole drug effect related
After the sample for determining the UAP1 expression level is collected by adopting the method in B, extracting total RNA of cells, determining the concentration by using Nanodrop, detecting and identifying the integrity by using agarose electrophoresis special for RNA, synthesizing cDNA by adopting the method in C, and finally determining the UAP1 expression level by adopting the method in D by using real-time fluorescence quantitative PCR.
FIG. 2 shows the relationship between the expression level of rabeprazole candidate drug and UAP1, wherein the expression level of UAP1 can be reduced by adding rabeprazole into cells, and the efficacy of rabeprazole is that the expression level of UAP1 is reduced to influence the hexosamine metabolic pathway (HBP) so as to exert the anti-tumor effect.
Example 3: effect of rabeprazole on the Activity of multiple myeloma cells NCI-H929
The experimental method for measuring cell viability by using the CCK8 assay described above, wherein NCI-H929 cells were used as 10 in the experimental group and the control wells 3 -10 4 Density of individual cells/well were cultured in 96 well plates in 100. Mu.L medium, and various concentrations of rabeprazole were added to the wells of the experimental group to form 10nM and 10. Mu.M of rabeprazole, and the cells were further cultured in an incubator for 48 hours. Adding CCK8 toAfter incubation for 1-4 hours in the incubator, the absorbance was measured at 450nm after gentle mixing on an orbital shaker to ensure uniform color distribution.
Cell viability was calculated: cell viability (%) = [ (As-Ab)/(Ac-Ab) ]x100%;
where as=experimental well absorbance (absorbance of wells containing cells, medium, CCK-8 and drug to be tested);
ab = blank well absorbance (absorbance of wells containing medium and CCK-8);
ac = control well absorbance (absorbance of wells containing cells, medium and CCK-8);
as shown in FIG. 3, the concentration of rabeprazole at 10nM can inhibit NCI-H929, and the cell viability can be reduced to about 50% at 10. Mu.M.
Example 4: effect of rabeprazole on the Activity of multiple myeloma cells RPMI-8226
The experimental method for measuring the cell viability by adopting the CCK8 assay is adopted, wherein RPMI-8226 cells are used for 10 in the experimental group and the control hole 3 -10 4 Density of individual cells/well were cultured in 96 well plates in 100. Mu.L of medium, rabeprazole at different concentrations was added to the experimental group wells to form a 10. Mu.M experimental group of rabeprazole, and the cells were further cultured in an incubator for 48 hours. After adding CCK8 and incubating in the incubator for 1-4 hours, the absorbance was measured at 450nm after gentle mixing on an orbital shaker to ensure uniform color distribution.
Cell viability was calculated according to the calculation formula of example 3.
The calculation results are shown in FIG. 4, and the rabeprazole also shows good inhibition effect on the RPMI-8226 of the multiple myeloma cells at the concentration of 10 mu M.
Example 5: effect of rabeprazole on ARD Activity of multiple myeloma cells
The experimental method for measuring cell viability by using the CCK8 assay described above, wherein ARD cells were used at 10 in the experimental group and control wells 3 -10 4 Density of individual cells/well in 96 well plates in 100. Mu.L medium, rabeprazole was added at different concentrationsIn the experimental group wells, a 10. Mu.M experimental group of rabeprazole was formed, and the cells were further cultured in an incubator for 48 hours. After adding CCK8 and incubating in the incubator for 1-4 hours, the absorbance was measured at 450nm after gentle mixing on an orbital shaker to ensure uniform color distribution.
Cell viability was calculated according to the calculation formula of example 3.
The results of the calculation are shown in fig. 5, and rabeprazole also shows good inhibition on multiple myeloma cell ARD at a concentration of 10 μm.
Example 6: effect of rabeprazole and bortezomib combination on the Activity of multiple myeloma cells NCI-H929
The experimental method for measuring cell viability by using the CCK8 assay described above, wherein NCI-H929 cells were used as 10 in the experimental group and the control wells 3 -10 4 The density of individual cells/well was cultured in 100 μl of medium in 96 well plates, rabeprazole, bortezomib and a combination of rabeprazole and bortezomib were added to the wells of the different experimental groups, respectively, to form a 10 μm experimental group of rabeprazole, a 10nM experimental group of bortezomib, a 10 μm combined experimental group of rabeprazole and bortezomib, and the cells were further cultured in an incubator for 48 hours. After adding CCK8 and incubating in the incubator for 1-4 hours, the absorbance was measured at 450nm after gentle mixing on an orbital shaker to ensure uniform color distribution.
Cell viability was calculated according to the calculation formula of example 3.
As shown in FIG. 6, 10. Mu.M rabeprazole can reduce the cell viability to about 50%, and 10nM bortezomib can reduce the cell viability to about 10%; the combination of rabeprazole and bortezomib can further reduce the cell survival rate, which is of great significance for further clearing away tiny residual focus and preventing recurrence.
Example 7: antitumor effect of rabeprazole in cell NCI-H929 subcutaneous xenograft model
Adopting the test method of subcutaneous xenograft, wherein the test group comprises;
1. control (NC): NOG mice were not administered, and the same volume of physiological saline was administered;
2. bortezomib group (BTZ): NOG mice were given 1mg/kg (dose administered per kg body weight) of bortezomib;
3. rabeprazole group (RAB): NOG mice were given 1mg/kg (dose administered per kg body weight) of rabeprazole;
4. rabeprazole group (RAB): NOG mice were given 2.5mg/kg (dose administered per kg body weight) of rabeprazole;
5. rabeprazole group (RAB): NOG mice were given 5mg/kg (dose administered per kg body weight) of rabeprazole;
as shown in the experimental result in figure 7, the drug effect of the rabeprazole 5mg/kg group and the positive drug bortezomib 1mg/kg group is similar, and compared with the control group, the rabeprazole 5mg/kg group and the bortezomib 1mg/kg group can generate obvious anti-tumor effect.
Example 8: antitumor effect of rabeprazole and bortezomib combination in cell NCI-H929 subcutaneous xenograft model
Adopting the test method of subcutaneous xenograft, wherein the test group comprises;
1. control (NC): NOG mice were not administered, and the same volume of physiological saline was administered;
2. bortezomib group (BTZ): NOG mice were given 1mg/kg bortezomib;
3. rabeprazole group (RAB): NOG mice were given 5mg/kg of rabeprazole;
4. combination group (btz+rab); NOG mice were given 1mg/kg bortezomib and 5mg/kg rabeprazole.
As shown in the experimental result in figure 8, the drug effect of the rabeprazole 5mg/kg group and the positive drug bortezomib 1mg/kg group is similar, and compared with the control group, the rabeprazole 5mg/kg group and the bortezomib 1mg/kg group can generate obvious anti-tumor effect. The anti-tumor effect of the combined administration group is obviously better than that of the control group and slightly better than that of the single administration group.
Example 9: effect of omeprazole on the activity of multiple myeloma cells NCI-H929
The experimental method for measuring cell viability by using the CCK8 assay described above, wherein NCI-H929 cells were used as 10 in the experimental group and the control wells 3 -10 4 Density of individual cells/well in 96 well plates in 100. Mu.L medium, different concentrations were usedOmeprazole was added to the experimental group wells to form an omeprazole 10nM experimental group and a 10. Mu.M experimental group, and the cells were cultured in an incubator for additional 48 hours. After adding CCK8 and incubating in the incubator for 1-4 hours, the absorbance was measured at 450nm after gentle mixing on an orbital shaker to ensure uniform color distribution.
Cell viability was calculated according to the calculation formula in example 3.
The results of the calculation are shown in fig. 9, and the effect of omeprazole on cell viability is similar to that of rabeprazole.
Example 10: effect of pantoprazole on the Activity of multiple myeloma cells NCI-H929
The experimental method for measuring cell viability by using the CCK8 assay described above, wherein NCI-H929 cells were used as 10 in the experimental group and the control wells 3 -10 4 Individual cell/well densities were cultured in 96-well plates in 100 μl of medium, pantoprazole at different concentrations was added to the experimental group wells to form a pantoprazole 10nM experimental group and a 10 μm experimental group, and the cells were further cultured in an incubator for 48 hours. After adding CCK8 and incubating in the incubator for 1-4 hours, the absorbance was measured at 450nm after gentle mixing on an orbital shaker to ensure uniform color distribution.
Cell viability was calculated according to the calculation formula in example 3.
The results of the calculation are shown in fig. 10, and the effect of pantoprazole on cell viability is similar to that of rabeprazole.
Example 11: effect of ilaprazole on the Activity of multiple myeloma cells NCI-H929
The experimental method for measuring cell viability by using the CCK8 assay described above, wherein NCI-H929 cells were used as 10 in the experimental group and the control wells 3 -10 4 Density of individual cells/well cells were cultured in 100. Mu.L medium in 96 well plates, ilaprazole at different concentrations was added to the wells of the experimental group to form 10nM and 10. Mu.M experimental groups of ilaprazole, and the cells were further cultured in an incubator for 48 hours. After adding CCK8 and incubating in the incubator for 1-4 hours, the absorbance was measured at 450nm after gentle mixing on an orbital shaker to ensure uniform color distribution.
Cell viability was calculated according to the calculation formula in example 3.
The results of the calculation are shown in fig. 11, and the effect of ilaprazole on cell viability is similar to that of rabeprazole.
Example 12: effect of lansoprazole on the Activity of multiple myeloma cells NCI-H929
The experimental method for measuring cell viability by using the CCK8 assay described above, wherein NCI-H929 cells were used as 10 in the experimental group and the control wells 3 -10 4 Density of individual cells/well were cultured in 96 well plates in 100. Mu.L medium, and lansoprazole at different concentrations was added to the wells of the experimental group to form 10nM and 10. Mu.M of lansoprazole experimental group, and the cells were further cultured in an incubator for 48 hours. After adding CCK8 and incubating in the incubator for 1-4 hours, the absorbance was measured at 450nm after gentle mixing on an orbital shaker to ensure uniform color distribution.
Cell viability was calculated according to the calculation formula in example 3.
The results of the calculation are shown in fig. 12, and the effect of lansoprazole on cell viability is similar to that of rabeprazole.
Example 13: influence of Esomeprazole on the Activity of multiple myeloma cells NCI-H929
The experimental method for measuring cell viability by using the CCK8 assay described above, wherein NCI-H929 cells were used as 10 in the experimental group and the control wells 3 -10 4 Density of individual cells/well were cultured in 96 well plates in 100. Mu.L of medium, and esomeprazole at different concentrations was added to the wells of the experimental group to form 10nM and 10. Mu.M of esomeprazole, and the cells were further cultured in an incubator for 48 hours. After adding CCK8 and incubating in the incubator for 1-4 hours, the absorbance was measured at 450nm after gentle mixing on an orbital shaker to ensure uniform color distribution.
Cell viability was calculated according to the calculation formula in example 3.
The results of the calculation are shown in fig. 13, and the effect of esomeprazole on cell viability is similar to that of rabeprazole.
Experiments prove that the rabeprazole, the omeprazole, the pantoprazole, the ilaprazole, the lansoprazole, the dexlansoprazole, the tenatoprazole, the leminoprazole, the Sha Weila oxazole, the picoprazole, the temozolomide and the esomeprazole can inhibit the activity of multiple myeloma cells at a low concentration on a cellular level, the rabeprazole can inhibit the activity of the multiple myeloma cells at an animal level at a dosage not exceeding a clinical dosage, and the proton pump inhibitors can inhibit the multiple myeloma cells at the cellular level and the animal level based on the proton pump inhibitors, so that the rabeprazole can be used for preparing medicaments for treating the multiple myeloma.
Although embodiments of the present invention have been disclosed above, it is not limited to the details and embodiments shown and described, it is well suited to various fields of use for which the invention would be readily apparent to those skilled in the art, and accordingly, the invention is not limited to the specific details and illustrations shown and described herein, without departing from the general concepts defined in the claims and their equivalents.

Claims (7)

1. The novel application of the proton pump inhibitor is characterized in that the proton pump inhibitor is applied to the preparation of medicines for treating multiple myeloma.
2. The novel use of the proton pump inhibitor according to claim 1, wherein the proton pump inhibitor comprises rabeprazole, omeprazole, pantoprazole, ilaprazole, lansoprazole, dexlansoprazole, tenatoprazole, leminoprazole, sha Weila oxazole, picoprazole, temozolomide and esomeprazole, and a compound preparation containing the above-mentioned corresponding proton pump inhibitor active ingredients.
3. The novel use of a proton pump inhibitor as claimed in claim 1, wherein the proton pump inhibitor is rabeprazole and the use of rabeprazole in combination with bortezomib for the preparation of a medicament for the treatment of multiple myeloma.
4. The novel use of a proton pump inhibitor as claimed in any one of claims 1 to 3, wherein the medicament for treating multiple myeloma further comprises a pharmaceutically acceptable carrier or pharmaceutical adjuvant for the proton pump inhibitor.
5. The novel use of a proton pump inhibitor as claimed in any one of claims 1 to 3, wherein the dosage form of the medicament for treating multiple myeloma comprises an oral preparation and an injection preparation.
6. The novel use of a proton pump inhibitor as claimed in claim 5, wherein the oral formulation comprises: tablets, capsules, oral liquids, buccal tablets, granules, suspension, dripping pills, sustained release agents, controlled release agents and targeting preparations.
7. The novel use of a proton pump inhibitor as claimed in claim 5, wherein the injectable formulation comprises: injection and powder injection.
CN202310419576.0A 2023-03-13 2023-04-19 New use of proton pump inhibitor Pending CN116603063A (en)

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