CN112138066B

CN112138066B - Application of black nightshade or black nightshade extract in increasing antibiotic or antineoplastic drug sensitivity

Info

Publication number: CN112138066B
Application number: CN202010938422.9A
Authority: CN
Inventors: 李岱; 李湘平; 付喆; 魏康竞; 付四海
Original assignee: Xiangya Hospital of Central South University
Current assignee: Xiangya Hospital of Central South University
Priority date: 2020-09-09
Filing date: 2020-09-09
Publication date: 2022-02-25
Anticipated expiration: 2040-09-09
Also published as: CN112138066A

Abstract

The invention relates to an application of black nightshade or black nightshade extract in increasing the sensitivity of antibiotics or antitumor drugs, belonging to the technical field of medicines. The research of the invention finds that the black nightshade or black nightshade extract is used in combination with antibiotics or antitumor drugs, so that the sensitivity of the antibiotics or antitumor drugs can be increased, the dosage can be reduced, the safety of the drug use can be improved, and the treatment time can be prolonged. The pharmaceutical composition provided by the invention can intervene in DNA abnormal methylation and histone modification mechanism based on the black nightshade or black nightshade extract, and can be used for treating antibiotic or tumor drug resistance. The invention provides a new solution for the drug resistance of the existing antibiotics or antitumor drugs, fully utilizes and prolongs the life cycle of the existing drugs, and is more in line with the current pharmaceutical industry level and the current medical investment situation in principle from the medical economy.

Description

Application of black nightshade or black nightshade extract in increasing antibiotic or antineoplastic drug sensitivity

Technical Field

The invention relates to an application of black nightshade or black nightshade extract in increasing the sensitivity of antibiotics or antitumor drugs, belonging to the technical field of medicines.

Background

Antibiotics and antitumor drugs are the main means for treating infectious diseases and tumors, respectively, however, the problem of drug resistance of bacteria and tumors is getting more serious due to the evolution mechanism of bacteria and tumors themselves and the long-term large dose use of antibiotics and antitumor drugs.

The natural medicine contains rich bioactive substances, and a large number of researches show that the traditional Chinese medicine and the western medicine can be combined in a synergistic manner through scientific compatibility, so that the sensitivity is increased, the dosage is reduced, and the toxic and side effects are reduced. The traditional Chinese medicine is used together with antibiotics and antitumor drugs, and the special advantages in the aspect of improving the drug resistance of bacteria and tumors are possibly played.

The black nightshade has the effects of clearing away heat and toxic materials, promoting blood circulation, removing blood stasis, inducing diuresis, relieving swelling, relieving cough and eliminating phlegm. Is clinically used for treating various inflammations and tumors, and the anti-tumor effect of the traditional Chinese medicine is a research hotspot of scholars at home and abroad. The combination of solanine and Doxorubicin (DOX) is also found to obviously improve the killing effect of DOX on leukemia K562/ADR cells, reverse the multidrug resistance of cells, and the mechanism of the combination is related to the inhibition of JNK activity, the down-regulation of MRP1 protein expression and the increase of accumulation of DOX in cells.

Epigenetic and protein modifications play an important role in bacterial and tumor resistance, while traditional Chinese medicines can interfere with DNA abnormal methylation and histone modification. Epigenetics mainly includes DNA methylation, histone modification, chromatin remodeling, post-transcriptional regulation, etc., and post-translational protein modification refers to chemical modification by adding different chemical groups to amino acid residues of proteins, both of which largely expand the coding capacity of bacterial and tumor cell genomes and promote the formation of drug resistance. Hsieh HY and the like carry out systematic epigenetic study on 48491 compounds in 3294 traditional Chinese medicines, and the result shows that 29.8 percent of traditional Chinese medicines or compound medicines can regulate and control functions of epigenome and miRNA. The solanum nigrum and the extract thereof have the effects of intervening the abnormal methylation of DNA and histone modification of bacteria and tumor cells, but no research on the combination treatment of bacteria and tumor resistance by the solanum nigrum, antibiotics and antitumor drugs based on the regulation and control mechanism of epigenetic inheritance exists at present.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides the application of the black nightshade or the black nightshade extract in increasing the sensitivity of antibiotics or antitumor drugs.

In order to achieve the purpose, the invention adopts the technical scheme that: the application of Solanum nigrum or Solanum nigrum extract in increasing antibiotic or antitumor drug sensitivity is provided.

The research of the invention finds that the black nightshade or the black nightshade extract can interfere DNA abnormal methylation and histone modification, can be combined with antibiotics or antitumor drugs, and can be used for treating bacterial or tumor drug resistance, increasing drug sensitivity, reducing dosage and prolonging treatment time. Experiments prove that the invention uses the black nightshade or black nightshade extract together with antibiotics or antitumor drugs, the black nightshade can intervene abnormal methylation of DNA and histone modification of bacteria and tumor cells, enhance the sensitivity of the drugs, improve indexes such as minimum inhibitory concentration, tumor inhibition rate and the like, improve the curative effect and provide a new scheme for solving the problem of drug resistance of bacteria and tumors in clinic.

As a preferred embodiment of the application of the invention, the black nightshade or black nightshade extract intervenes DNA abnormal methylation and histone modification, increasing the sensitivity of antibiotics or antitumor drugs.

In a second aspect, the present invention provides a pharmaceutical composition for increasing sensitivity to an antibiotic or an anti-tumor drug, the pharmaceutical composition comprising an antibiotic or an anti-tumor drug, and Solanum nigrum or an extract of Solanum nigrum.

As a preferred embodiment of the pharmaceutical composition of the present invention, the black nightshade is a fresh black nightshade fruit, and the black nightshade extract is solanine.

As a preferable embodiment of the pharmaceutical composition of the present invention, the anti-tumor drug is an anti-tumor biological drug or an anti-tumor chemical drug.

In a preferred embodiment of the pharmaceutical composition of the present invention, the anti-tumor biological agent is nivolumab and the anti-tumor chemical agent is gefitinib.

In a preferred embodiment of the pharmaceutical composition of the present invention, the mass ratio of the black nightshade or black nightshade extract to the antibiotic is (50: 1) - (1: 1), the mass ratio of the black nightshade or black nightshade extract to the anti-tumor biological drug is (50: 1) - (1: 1), and the mass ratio of the black nightshade or black nightshade extract to the anti-tumor chemical drug is (100: 1) - (1: 1).

In a preferred embodiment of the pharmaceutical composition of the present invention, the mass ratio of the black nightshade or black nightshade extract to the antibiotic is (30: 1) - (5: 1), the mass ratio of the black nightshade or black nightshade extract to the anti-tumor biological drug is (20: 1) - (5: 1), and the mass ratio of the black nightshade or black nightshade extract to the anti-tumor chemical drug is (30: 1) - (10: 1).

In a third aspect, the present invention provides a preparation method of the above pharmaceutical composition, comprising the following steps:

(1) crushing Solanum nigrum fruit into coarse powder, adding 6 times of water, stirring and extracting at 20 ℃ for 0.5-4 hours for 2 times, monitoring the content of solanine in an extracting solution by adopting a high performance liquid chromatography in the extraction process, ensuring that the content of the solanine in the obtained freeze-dried powder is 3-6mg/g, stopping extraction when the corresponding content is reached, combining the extracting solutions, filtering, centrifuging, filling a centrifugate into a freeze-drying box, freeze-drying, extracting and grinding to obtain fine powder which is sieved by a 200-mesh sieve for later use;

or grinding the Solanum nigrum extract and sieving with a 200-mesh sieve to obtain fine powder for later use;

(2) placing antibiotic or antitumor drug into a stirrer, stirring uniformly, extracting and grinding to obtain fine powder which is sieved by a 200-mesh sieve for later use;

(3) and (3) mixing the fine powder obtained in the steps (1) to (2), uniformly stirring, putting into an oven, drying at 40 ℃, cooling, extracting, grinding, and sieving with a 200-mesh sieve to obtain the pharmaceutical composition.

In a fourth aspect, the invention provides an oral tablet or capsule, which comprises the pharmaceutical composition and pharmaceutically acceptable auxiliary materials. The preparation method of the oral tablet or capsule comprises the following steps: mixing the medicinal composition and pharmaceutically acceptable adjuvants uniformly.

Compared with the prior art, the invention has the beneficial effects that: the research of the invention finds that the black nightshade or black nightshade extract is used in combination with antibiotics or antitumor drugs, so that the sensitivity of the antibiotics or antitumor drugs can be increased, the dosage is reduced, and the treatment time is prolonged. The pharmaceutical composition provided by the invention can intervene in DNA abnormal methylation and histone modification mechanism based on the black nightshade or black nightshade extract, and can be used for treating antibiotic or tumor drug resistance. The invention provides a new solution for the drug resistance of the existing antibiotics or antitumor drugs, fully utilizes and prolongs the life cycle of the existing drugs, and is more in line with the current pharmaceutical industry level and the current medical investment situation in principle from the medical economy.

Drawings

FIG. 1 is a photograph showing the tumor-bearing mice of the effect example 6 after they died, wherein (a) from left to right are photographs showing the tumor-bearing mice of examples 22 to 26 after they died, respectively, and (b) from left to right are photographs showing the tumor-bearing mice of comparative examples 7 to 8 after they died, respectively.

Detailed Description

To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to specific examples.

Example 1

A pharmaceutical composition for increasing antibiotic drug sensitivity comprises an antibiotic and black nightshade, wherein the mass ratio of the antibiotic to the black nightshade and the type of the antibiotic in the pharmaceutical composition are shown in Table 1.

The preparation method of the pharmaceutical composition of this example comprises:

(2) putting the antibiotics into a stirrer, stirring uniformly, extracting and grinding to obtain fine powder which is sieved by a 200-mesh sieve for later use;

Examples 2 to 7 and comparative examples 1 to 2

The pharmaceutical compositions of examples 2-7 comprise antibiotics and black nightshade, and the mass ratio and the type of the antibiotics in the pharmaceutical compositions are shown in table 1. Comparative example 1 contained no solanum nigrum and comparative example 2 contained no antibiotics.

The preparation methods of the pharmaceutical compositions of examples 2-7 and comparative examples 1-2 are the same as example 1.

TABLE 1

Examples 8 to 14 and comparative examples 3 to 4

The pharmaceutical composition of examples 8 to 14 includes the antitumor drug and nightshade, and the mass ratio and the type of the antitumor drug of the pharmaceutical composition are shown in table 2. Comparative example 3 did not contain solanum nigrum and comparative example 4 did not contain anti-tumor biopharmaceuticals.

The preparation methods of the pharmaceutical compositions of examples 8 to 14 and comparative examples 3 to 4 are the same as example 1.

TABLE 2

	Anti-tumor biological medicine	The mass ratio of the black nightshade to the anti-tumor biological medicine
			Example 8	Nivolumab	50:1
Example 9	Nivolumab	20:1
			Example 10	Nivolumab	10:1
Examples11	Nivolumab	5:1
			Example 12	Nivolumab	1:1
Example 13	Bevacizumab	10:1
			Example 14	Adalimumab	10:1
Comparative example 3	Nivolumab	Contains no black nightshade
			Comparative example 4	--	Contains no antitumor biological medicine

Examples 15 to 21 and comparative examples 5 to 6

The pharmaceutical composition of examples 15 to 21 includes an antitumor chemical and solanum nigrum, and the mass ratio and the type of the antitumor chemical of the pharmaceutical composition are shown in table 3. Comparative example 5 did not contain solanum nigrum and comparative example 6 did not contain anti-tumor chemicals.

The preparation methods of the pharmaceutical compositions of examples 15 to 21 and comparative examples 5 to 6 are the same as example 1.

TABLE 3

	Anti-tumor chemical	The mass ratio of the black nightshade to the anti-tumor chemical medicine
			Example 15	Gefitinib	100:1
Example 16	Gefitinib	30:1
			Example 17	Gefitinib	20:1
Example 18	Gefitinib	10:1
			Example 19	Gefitinib	1:1
Example 20	5-Fluorouracil	20:1
			Example 21	Paclitaxel	20:1
Comparative example 5	Gefitinib	Contains no black nightshade
			Comparative example 6	--	Contains no antitumor chemical

Examples 22 to 26 and comparative examples 7 to 8

The pharmaceutical compositions of examples 22 to 26 include the antitumor chemical and solanum nigrum, and the mass ratio and the type of the antitumor chemical of the pharmaceutical compositions are shown in table 4. Comparative example 7 contained no nightshade and comparative example 8 contained no anti-tumor chemicals.

The preparation methods of the pharmaceutical compositions of examples 22-26 and comparative examples 7-8 are the same as example 1.

TABLE 4

Effect example 1 in vitro antibacterial test

Drug-resistant strains such as escherichia coli, klebsiella pneumoniae, pseudomonas aeruginosa, acinetobacter baumannii and the like are collected, a 96-well plate trace broth two-fold dilution method is adopted, the concentration multiple dilution of the antibacterial drug is 0.063-128mg/ml, the total concentration is 12, and the use amount of the black nightshade is respectively tested by a fixed dose method and a method of equivalent multiple dilution with the antibacterial drug. Several colonies, freshly grown overnight, were picked, adjusted to 0.5 McLeod concentration in MH broth by turbidimetry, and after 20h of incubation at 35 ℃ the Minimum Inhibitory Concentration (MIC) was read and the corresponding quality control standard strain was used as a control for each experiment.

The results of the above tests on the pharmaceutical compositions of examples 1 to 7 and comparative examples 1 to 2 are shown in Table 5.

TABLE 5

According to test results, the pharmaceutical composition disclosed by the invention in the embodiments 1-7 has a lower minimum inhibitory concentration; comparative example 1 contains no black nightshade, and has a higher minimum inhibitory concentration; comparative example 2 contains no antibiotic and has a higher minimum inhibitory concentration than the present invention.

In addition, in the pharmaceutical compositions of examples 2 to 4, when the mass ratio of the solanum nigrum to the antibiotic is (30: 1) to (5: 1), the minimum inhibitory concentration is lower.

Effect example 2 in vivo antibacterial experiment

Collecting clinically isolated penicillin-insensitive Streptococcus pneumoniae strain prepared by adding 5% defibrinated sheep blood into blood culture medium (MH culture medium) at 35 deg.C and 5% CO₂Environment (CO)₂Incubator) for 16 h.

Establishing a mouse immune suppression model: reference method, mice were given intraperitoneal injections of 150, 100mg/kg Cyclophosphamide (CPM) on test day 1, day 4, respectively, to establish an immunosuppressive model.

Establishing a mouse streptococcus pneumoniae pneumonia model: single colony Streptococcus pneumoniae was inoculated into 10mL brain-heart attack solution with 5% CO₂Culturing at 35 deg.C for 8-12 h. Diluting with sterile brain and heart invasion solution, collecting 500 μ L, adding 500 μ L0.2% agar, shaking, mixing to obtain Streptococcus pneumoniae bacterial suspension with final concentration of about 1 × 107CFU/mL, and standing at 42-45 deg.C.

The last day after the mice were injected with cyclophosphamide intraperitoneally, i.e., day 5 of the experiment, streptococcus pneumoniae bacterial suspension was instilled through the nasal cavity on a sterile operating table to carry out streptococcus pneumoniae infection. Before bacterial inoculation, 0.12mL of 1.0% sodium pentobarbital is intraperitoneally injected for anesthesia, when the whole muscle of the mouse is relaxed, the mouse is converted into an anesthesia state from excitation, four limbs are fixed on a fixer, the fixer is erected to enable the mouse to be upright, and incisors are fixed by thin lines. Sucking 50 μ L of the suspension of Streptococcus pneumoniae bacteria with a micro liquid feeder, slowly dripping from nasal cavity, and standing for 5min after dripping to allow the bacteria liquid to be fully sucked. After 5min of infection, the mice were released and returned to their cages for waking.

Examples 1-7 in vivo antibacterial action against mouse pneumonia

The infected mice were divided into 9 groups of 5 mice each, and the drugs of examples 1 to 7 and comparative examples 1 to 2 were administered, respectively. A blank control group of 7 was set and given physiological saline.

After 2h of infection, 2 mice in the blank control group were sacrificed, bilateral lung tissues were immediately isolated under aseptic conditions, weighed and placed in 1.0mL of sterile physiological saline, homogenized in a tissue disperser, and counted for colonies. While other mice were given drug/saline treatment. Each group was administered at a dose of 234mg/kg (equivalent to 1 g/day in humans) in 2 divided doses of 0.2ml each at 12 hour intervals.

24 hours after the first administration (i.e. 26 hours after infection), all mice were sacrificed, lung tissue was dissected and taken, weighed, homogenized, and bacteria were cultured and counted to calculate the bacteria content per gram of lung tissue. Statistical differences between drugs were compared using SPSS software for two independent sample t-tests. The results are shown in Table 6.

TABLE 6

According to test results, the pharmaceutical composition disclosed by the invention in the embodiments 1-7 has a strong in-vivo antibacterial effect; comparative example 1 contains no Solanum nigrum and has a low in vivo antibacterial effect; comparative example 2 contains no antibiotic and its in vivo antibacterial effect is inferior to the present invention.

In addition, in the pharmaceutical compositions of examples 2 to 4, when the mass ratio of the solanum nigrum to the antibiotic is (30: 1) to (5: 1), the in vivo antibacterial effect is stronger.

Effect example 3 inhibition Effect test on proliferation of human Lung cancer cell line A549

By MTT method, will pairThe human lung cancer A549 cells in several growth stages are prepared into the culture medium with the density of 5 multiplied by 10⁴After the cell suspension was suspended in a volume of 10. mu.L/well in a 96-well plate, the culture medium was discarded after 24 hours of incubation. Each well was filled with 90. mu.L of serum-free medium, 10. mu.L of each of the solutions of group B of the above preparation examples was added to the administered group, and an equal volume of PBS solution was administered to the blank group. After continuing culturing for 36h, adding 10 mu L of 5mg/mL MTT solution into each well, incubating for 4h, stopping incubation, discarding the supernatant, adding 100 mu L of DMSO into each well, oscillating for 10min in a micro oscillator in a dark place, measuring the absorbance (A) value of each well by using an enzyme labeling instrument, measuring the absorbance (A) value for 6 times in parallel at a detection wavelength of 550nm and a reference wavelength of 690nm, and taking an average value. The inhibition rate of each of the solutions to be administered on the growth of A549 cells was calculated according to the following formula, and the half inhibitory concentration (IC50) was determined using SPSS software.

The cell growth inhibition rate was (1-mean a value of experimental group/mean a value of blank control group) × 100%.

The pharmaceutical compositions of examples 8 to 14, comparative examples 3 to 4, examples 15 to 21, and comparative examples 5 to 6 were subjected to the above-described tests. The results are shown in Table 7.

TABLE 7

Group of	IC50(nmol/L)
		Example 8	5989.2
Example 9	3908.1
		Example 10	3344.1
Example 11	4756.5
		Example 12	6942.2
Example 13	8327.9
		Example 14	8857.5
Comparative example 3	6405.0
		Comparative example 4	13118.9
Example 15	20.0
		Example 16	5.3
Example 17	5.5
		Example 18	16.0
Example 19	100.7
		Example 20	288.4
Example 21	1005.8
		Comparative example 5	20.1
Comparative example 6	13050.6

The test results show that the IC50 values of the pharmaceutical compositions of the invention in examples 8-14 and 15-21 are low; comparative example 3 and comparative example 5, which did not contain solanum nigrum, had higher IC50 values; comparative examples 4 and 6, which do not contain an anti-tumor chemical, have IC50 values higher than the present invention.

In addition, in the pharmaceutical compositions of examples 9 to 11, the mass ratio of the black nightshade to the antitumor biological agent is (20: 1) - (5: 1), and the IC50 value is lower; in the pharmaceutical compositions of examples 16 to 18, the weight ratio of the Solanum nigrum to the antitumor chemical is (30: 1) to (10: 1), and the IC50 value is lower.

Effect example 4 test for inhibition of proliferation of human Hodgkin lymphoma cell line L428

Using the MTT method, L428 cell line was cultured in DMEM medium containing 10% inactivated newborn calf serum, 100U/mL penicillin and 100. mu.g/mL streptomycin at 37 ℃ under 5% CO₂Culturing under the condition, and taking cells in logarithmic growth phase for experiment. L428 and 90. mu.L of BJAB cell line were cultured in 96-well plates at a concentration of 2X 106/ml, 10. mu.L of LPBS was added to the blank control group, and 10. mu.L of the liquid medicine of the above preparation examples was added to the administration group, respectively, to give the final concentration of the drug (calculated as crude drug). A negative control group is prepared by adding different dosages of liquid medicine into the culture solution without cells. Setting 6 parallel wells, culturing cells for 24h after administration, adding 20 μ L of 5.0mg/mL MTT solution into each well, shaking and mixing uniformly, placing in an incubator for continuous incubation for 4h, discarding supernatant in each well, adding 100 μ L of LDMSO to stop reaction, shaking and mixing uniformly for 10min, after purple crystals in each well are completely dissolved, measuring the absorbance (A) at 490nm by using a calibrator, and calculating the half inhibitory concentration (IC50) by using SPSS software.

The pharmaceutical compositions of examples 8 to 14, comparative examples 3 to 4, examples 15 to 21, and comparative examples 5 to 6 were subjected to the above-described tests. The results are shown in Table 8.

TABLE 8

Effect example 5 tumor suppression Rate test (lymphoma) for tumor-bearing mice

Tumor mouse model establishment: resuscitating mouse A20 lymphoma cells, adjusting concentration to 1 × 10⁸The cell suspension of each/mL is inoculated in the axilla of the right forelimb of a BALB/c mouse, and subcutaneous tumor nodules with the diameter of about 0.5cm can be observed for 1 week, which indicates that the establishment of a tumor mouse model is successful.

Grouping and administration treatment: mice were divided into 18 groups of 10 mice depending on the treatment regimen and the experiment was started one week after tumor inoculation.

Tumor volumes were measured with a vernier caliper at 1, 3, 5, 7, 9, 11 d. 24h after the last administration, the orbital bleeding is measured for blood routine. After sacrifice, weights of spleen, thymus and tumor were measured, and organ index and tumor suppression rate were calculated.

The pharmaceutical compositions of examples 8 to 14, comparative examples 3 to 4, examples 15 to 21, and comparative examples 5 to 6 were subjected to the above-described tests. The results are shown in Table 9.

TABLE 9

The test results show that the tumor inhibition rates of the pharmaceutical compositions of the invention in examples 8-14 and 15-21 are high; comparative examples 3 and 5, which do not contain solanum nigrum, have a lower tumor inhibition than the present invention; comparative examples 4 and 6, which do not contain an antitumor chemical, have a lower tumor inhibition rate than the present invention.

In addition, in the pharmaceutical compositions of examples 9 to 11, the weight ratio of the solanum nigrum to the anti-tumor biological agent is (20: 1) - (5: 1), and the tumor inhibition rate is higher; in the pharmaceutical compositions of examples 16 to 18, the ratio by mass of solanum nigrum to the anti-tumor chemical is (30: 1) to (10: 1), and the anti-tumor rate is higher.

Effect example 6 drug effect experiment in tumor-bearing mice (Lung cancer)

The mice were revived and 10 mice were randomly selected as blank control. The right axilla of the remaining mice were inoculated subcutaneously with Lewis cells (1X 10)⁷mL), 0.2mL each, after 24h, were randomly divided into 7 groups of 10. The tested groups are respectively and regularly administrated with the medicaments of examples 22 to 28 and comparative examples 7 to 8 by intragastric administration for 1 time per day and are continuously administrated for 42 days.

24h after the last administration, the orbital bleeding is measured for blood routine. After sacrifice, the liver, kidney, and tumor weights were weighed, and the organ index was calculated, and the results are shown in table 10 and fig. 1.

Watch 10

According to the test results, the survival rate of the pharmaceutical composition of the embodiment 22-26 is higher; comparative example 7 does not contain solanum nigrum, and the survival rate is lower than that of the present invention; comparative example 8, which does not contain an anti-tumor chemical, has a lower survival rate than the present invention. Based on the tumor weight data analysis, the example tumor/body weight ratios were less than the comparative examples, indicating that the combination of chemicals with solanum nigrum inhibits tumor growth. According to the analysis of the organ coefficient data, the difference between each group in the examples and the comparative example is not significant, and the black nightshade has no obvious liver and kidney toxicity.

In the pharmaceutical compositions of examples 23 to 25, the ratio by mass of solanum nigrum to the antitumor agent is (20: 1) to (5: 1), and the survival rate is higher.

Effect example 7 analysis of DNA methylase Activity

The effect of various doses of solanine on the activity of M.AluI methylase was examined, and no drug treatment was used as a control. 10 μ L of methylase protection reaction system (1 μ L of lambda DNA; 1 μ L of drug; 1 μ L of LSAM; 1 μ L of Alu I; 1 μ L of buffer solution; 5 μ L of LH2O), placing in a 37 ℃ water bath, after 2h, carrying out Alu I enzyme digestion reaction, 20 μ L of reaction system (5 μ L of the methylase cleavage protection reaction system; 2 μ L of buffer solution, 1 μ L of LAlu I, 12 μ L of LH2O), 37 ℃ for 2h, and 0.7% gel electrophoresis.

The test result shows that the lambda DNA is cut into a plurality of strips by the M.AluI enzyme compared with the control group in the solanine treatment group, which shows that the solanine influences the activity of the M.AluI enzyme, so that the lambda DNA can not be modified by the M.AluI enzyme, and thus the lambda DNA is cut by the enzyme.

Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. The application of the solanum nigrum extract in the preparation of drugs for increasing the sensitivity of antibiotics or antitumor drugs, wherein the antitumor drugs are nivolumab or gefitinib; the antibiotic is cefoperazone sulbactam;

the content of the solanine in the solanum nigrum extract is 3-6 mg/g.

2. The use of claim 1, wherein the tumor is lung cancer or lymphoma.

3. The use of claim 2, wherein the lymphoma is hodgkin's lymphoma.

4. The use of claim 1, wherein the antibiotic is used to inhibit or eliminate infection by escherichia coli, klebsiella pneumoniae, pseudomonas aeruginosa, acinetobacter baumannii, or streptococcus pneumoniae.

5. A pharmaceutical composition for increasing the sensitivity of antibiotics or antitumor drugs is characterized by comprising the antibiotics or antitumor drugs and a black nightshade extract; wherein the anti-tumor drug is nivolumab or gefitinib; the antibiotic is cefoperazone sulbactam; the content of the solanine in the solanum nigrum extract is 3-6 mg/g;

the mass ratio of the black nightshade extract to the antibiotics is (30: 1) - (5: 1), the mass ratio of the black nightshade extract to the nivolumab is (50: 1) - (1: 1), and the mass ratio of the black nightshade extract to the gefitinib is (100: 1) - (1: 1).

6. The pharmaceutical composition according to claim 5, wherein the mass ratio of the Solanum nigrum extract to nivolumab is (20: 1) - (5: 1), and the mass ratio of the Solanum nigrum extract to gefitinib is (30: 1) - (10: 1).

7. A process for the preparation of a pharmaceutical composition according to any one of claims 5 to 6, comprising the steps of:

(1) crushing solanum nigrum fruits into coarse powder, adding 6 times of water, stirring and extracting for 0.5-4 hours at 20 ℃, extracting for 2 times, monitoring the content of solanine in an extracting solution by adopting a high performance liquid chromatography in the extracting process, ensuring that the content of the solanine in the obtained freeze-dried powder is 3-6mg/g, stopping extracting when the corresponding content is reached, combining the extracting solutions, filtering, centrifuging, filling a centrifugate into a freeze-drying box, freeze-drying, extracting and grinding to obtain fine powder which is sieved by a 200-mesh sieve for later use;

(3) and (3) mixing the fine powder obtained in the steps (1) to (2), stirring uniformly, putting into an oven, drying at 40 ℃, cooling, extracting, grinding, and sieving with a 200-mesh sieve to obtain the pharmaceutical composition.

8. An oral tablet or capsule, wherein the tablet or capsule comprises the pharmaceutical composition of any one of claims 5 to 6 and pharmaceutically acceptable excipients.