CN110279693B - Application of composition in preparation of medicine for preventing and/or treating fever with thrombocytopenia syndrome virus - Google Patents
Application of composition in preparation of medicine for preventing and/or treating fever with thrombocytopenia syndrome virus Download PDFInfo
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- CN110279693B CN110279693B CN201910679853.5A CN201910679853A CN110279693B CN 110279693 B CN110279693 B CN 110279693B CN 201910679853 A CN201910679853 A CN 201910679853A CN 110279693 B CN110279693 B CN 110279693B
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Abstract
The application relates to the field of medicines, in particular to application of a composition in preparing a medicine for preventing and/or treating fever with thrombocytopenia syndrome virus. The application of a composition in preparing a medicament for preventing and/or treating fever with thrombocytopenia syndrome virus, wherein the composition comprises nifedipine or pharmaceutically acceptable salts, solvates or hydrates thereof. The application provides a new application of nifedipine or pharmaceutically acceptable salts, solvates or hydrates thereof, which can be used for preparing medicines for preventing and/or treating fever with thrombocytopenia syndrome virus.
Description
Technical Field
The application relates to the field of medicines, in particular to application of a composition in preparing a medicine for preventing and/or treating fever with thrombocytopenia syndrome virus.
Background
SFTSV (SFTSV), the main symptoms caused by SFTSV infection include Fever, hemorrhage, gastrointestinal discomfort, muscle soreness, lymph node enlargement, Thrombocytopenia, leukopenia and the like; the virus is mainly carried and transmitted by ticks, tick bites are important reasons causing human infection, SFTSV can infect people of nearly all ages, and the virus has great threat to old people and has high fatality rate. SFTSV belongs to the order Bunyavirales (Bunyavirales), the family Phenguiviridae, the genus Phlebovirus (Phlebovirus). Is a minus-strand RNA virus with segmented genome, the genome is divided into three segments according to the length (L, M, S), and the segments are wrapped by Nucleocapsin (NP).
The prevention and treatment of viral diseases mainly depends on vaccines and drugs. Currently, there are few reports of vaccine development and drug development for SFTSV. The existing treatment methods aiming at the SFTSV infection are quite limited, the main treatment means are symptomatic support treatment and broad-spectrum antiviral treatment, the curative effect is limited, and the individual difference is large.
Disclosure of Invention
The embodiment of the application aims to provide application of a composition in preparing a medicine for preventing and/or treating fever with thrombocytopenia syndrome virus, and aims to provide a medicine capable of being used for preventing and/or treating SFTSV.
The application provides an application of a composition in preparing a medicine for preventing and/or treating fever with thrombocytopenia syndrome virus, wherein the composition comprises nifedipine or pharmaceutically acceptable salts, solvates or hydrates thereof.
Nifedipine has an anti-SFTSV effect and can be used for inhibiting replication of SFTSV. Nifedipine or a pharmaceutically acceptable salt, solvate or hydrate thereof can reduce mortality of mice caused by SFTSV infection. Nifedipine or a pharmaceutically acceptable salt, solvate or hydrate thereof can be used for preparing a medicament for preventing and/or treating fever with thrombocytopenia syndrome virus. The medicine can be used for treating SFTSV and expanding the application of nifedipine.
In some embodiments of the first aspect of the present application, the composition comprises 50 μ Mol/L to 400 μ Mol/L of said nifedipine or a pharmaceutically acceptable salt, solvate or hydrate thereof.
At this concentration, the dosage is safe and the anti-SFTSV effect is obvious.
In some embodiments of the first aspect of the present application, the composition comprises 120 μ Mol/L to 170 μ Mol/L of said nifedipine or a pharmaceutically acceptable salt, solvate or hydrate thereof.
In some embodiments of the first aspect of the present application, the composition comprises 150 μ Mol/L to 155 μ Mol/L of said nifedipine.
At this concentration, the dose is safe and the anti-SFTSV effect is good.
In some embodiments of the first aspect of the present application, the composition is for use in the preparation of a medicament for inhibiting replication of the fever with thrombocytopenia syndrome virus.
Nifedipine can inhibit the replication of SFTSV and therefore can be used for preparing SFTSV inhibitors.
In some embodiments of the first aspect of the present application, the composition further comprises a pharmaceutically acceptable adjuvant.
In some embodiments of the first aspect of the present application, the dosage form of the composition is a tablet.
In some embodiments of the first aspect of the present application, the dosage form of the drug is a gastrointestinal administration dosage form.
In some embodiments of the first aspect of the present application, the administration of the drug is oral.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are required to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present application and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained from the drawings without inventive effort.
Fig. 1 shows the test results of test example 1 of the present application.
Fig. 2 shows the test results of test example 2 of the present application.
Fig. 3 shows the test results of test example 3 of the present application.
Fig. 4 shows the test results of test example 4 of the present application.
Fig. 5 shows the test results of test example 5 of the present application.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions of the embodiments of the present application will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The following is a specific description of the use of the composition of the embodiments of the present application in the preparation of a medicament for the prevention and/or treatment of fever with thrombocytopenia syndrome virus.
The application of a composition in preparing a medicament for preventing and/or treating fever with thrombocytopenia syndrome virus is disclosed, wherein the composition comprises nifedipine or pharmaceutically acceptable salts, solvates or hydrates thereof.
Further, the composition comprising nifedipine or a pharmaceutically acceptable salt, solvate or hydrate thereof means that the composition comprises at least one of nifedipine, a pharmaceutically acceptable salt of nifedipine, a pharmaceutically acceptable solvate of nifedipine or a pharmaceutically acceptable hydrate of nifedipine.
In this embodiment, "prevention" refers to the possibility of reducing or eliminating the disease. "treating" or "treatment" refers to the complete or partial elimination or alleviation of the disease and/or its attendant symptoms.
"/" means an alternative. The medicine for preventing and/or treating fever with thrombocytopenia syndrome virus is: a medicament for preventing and treating fever with thrombocytopenia syndrome virus, or a medicament for preventing fever with thrombocytopenia syndrome virus, or a medicament for treating fever with thrombocytopenia syndrome virus.
Fever with Thrombocytopenia Syndrome (Severe Fever with Thrombocytopenia Syndrome, SFTS for short).
Nifedipine: 1, 4-dihydro-2, 6-dimethyl-4- (2-nitrophenyl) -3, 5-pyridinedicarboxylic acid dimethyl ester, the chinese alias: reserpine, XINTONGDING, nifedipine; the name of English: nifedipine, molecular formula: c17H18N2O6。
Structural formula (xvi):
nifedipine in pharmaceutically acceptable solvates, such as nifedipine-1, 4 dioxane solvate, nifedipine-morpholine solvate.
It should be noted that in the examples of the present application, nifedipine may be crystalline, amorphous, and the like, and accordingly, the crystalline form of nifedipine is not limited.
After research, the inventor finds that nifedipine has the effect of resisting SFTSV and can be used for inhibiting the replication of the SFTSV. Nifedipine or a pharmaceutically acceptable salt, solvate or hydrate thereof can reduce mortality of mice caused by SFTSV infection. Can effectively inhibit replication of fever with thrombocytopenia syndrome virus.
Further, the composition comprises 50-400 mu Mol/L of nifedipine or pharmaceutically acceptable salt, solvate or hydrate thereof. At this concentration, the dosage is safe and the anti-SFTSV effect is obvious.
The composition comprises 50-400 mu Mol/L of nifedipine or pharmaceutically acceptable salt, solvate or hydrate thereof; that is, the composition may comprise 50 to 400 μ Mol/L of nifedipine, or the composition may comprise 50 to 400 μ Mol/L of a pharmaceutically acceptable salt, solvate or hydrate of nifedipine.
Further, the concentration of nifedipine or its pharmaceutically acceptable salt, solvate or hydrate is 100-280. mu. Mol/L, and further 200-280. mu. Mol/L, such as 60. mu. Mol/L, 80. mu. Mol/L, 100. mu. Mol/L, 120. mu. Mol/L, 150. mu. Mol/L, 170. mu. Mol/L, 200. mu. Mol/L, 250. mu. Mol/L, 300. mu. Mol/L, etc.
Further, the concentration of nifedipine or pharmaceutically acceptable salts, solvates or hydrates thereof is 120-170 mu Mol/L. Further, the concentration of nifedipine or pharmaceutically acceptable salts, solvates or hydrates thereof is 150-. At this concentration, the dose is safe and the anti-SFTSV effect is good.
Further, the composition also comprises pharmaceutically acceptable auxiliary agents. The composition may be in the form of a solid or a liquid or a paste.
Further, the medicaments may be formulated with the compositions using pharmaceutically acceptable carriers and/or excipients as adjuvants, for preparation in unit dosage form, or for inclusion in multi-dose containers. The formulation may be a solution, suspension or emulsion (emulsified solution), extract, powder, granule, tablet or capsule in an oil or aqueous medium, and may further comprise a dispersing agent or a stabilizer.
Pharmaceutically acceptable adjuvants include, but are not limited to, fillers, binders, preservatives, disintegrants, sweeteners, release control agents, stabilizers and the like.
Illustratively, the filler may be selected from at least one of glucose, sucrose, maltose, starch, sorbitol, maltitol, xylitol, dextrin, maltodextrin cellulose or cellulose derivatives.
The binder may be selected from at least one of povidone, methylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, hydroxyethylcellulose, gelatin, gum, sucrose, or starch.
The preservative may be selected from at least one of benzoic acid, sodium benzoate, benzyl alcohol, butylated hydroxyanisole, butylated hydroxytoluene, chlorobutanol, gallic acid ester, hydroxybenzoate or EDTA.
The disintegrant may be selected from at least one of sodium starch glycolate, cross-linked polyvinylpyrrolidone, cross-linked carboxymethylcellulose, starch, or microcrystalline cellulose.
The sweetener may be at least one selected from sucralose, saccharin, sodium saccharin, potassium saccharin, calcium saccharin, potassium acesulfame or sodium cyclamate, mannitol, fructose, sucrose, or maltose.
In this embodiment, the dosage form of the drug is a dosage form for gastrointestinal administration, such as oral liquid, buccal tablet or tablet. Further, in this embodiment, the drug is a tablet, and the administration mode of the drug is oral.
It is noted that in other embodiments of the present application, the drug may be administered parenterally, which may include intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, endothelial administration, topical administration, intranasal administration, rectal administration, and the like.
In the case of oral administration, the composition may be administered by any device capable of transferring the active substance to the target cells. The route of administration may vary depending on the general condition and age of the subject to be treated, the nature of the treatment condition and the active ingredient selected.
In addition, the dosage form of the medicament may be administered in the form of suppositories, sprays, ointments, creams, gels, inhalants or skin patches.
The pharmaceutical compositions can also be prepared for administration to an animal, e.g., to a human.
Further, in the examples of the present application, the dosage of the drug may be adjusted depending on factors such as formulation method, administration method, age, body weight and sex of a patient, pathological conditions, diet, administration times, administration route, excretion rate and response sensitivity, and a physician having ordinary skill can easily determine and prescribe a dose effective for the desired treatment or prevention.
A preparation method of a medicine for preventing and/or treating fever with thrombocytopenia syndrome virus comprises the steps of mixing a composition and auxiliary materials, wherein the composition comprises nifedipine or pharmaceutically acceptable salts, solvates or hydrates thereof.
The application provided by the embodiment of the application can increase the application range of nifedipine or pharmaceutically acceptable salts, solvates or hydrates thereof, and has a new application of resisting SFTSV.
The features and properties of the present application are described in further detail below with reference to examples.
Example 1
The embodiment provides a medicament, which is prepared from a composition and auxiliary materials, wherein the composition comprises nifedipine. The concentration of nifedipine in the composition was 150. mu. Mol/L.
The medicine can be used for preventing and/or treating fever with thrombocytopenia syndrome virus.
Example 2
The embodiment provides a medicine which is prepared from a composition and auxiliary materials, wherein the composition comprises 170 mu Mol/L of nifedipine.
The medicine can be used for preventing and/or treating fever with thrombocytopenia syndrome virus. Accordingly, the above drugs can inhibit replication of fever with thrombocytopenia syndrome virus.
Example 3
The embodiment provides a medicine which is prepared from a composition and auxiliary materials, wherein the composition comprises 170 mu Mol/L of nifedipine.
The medicine can be used for preventing and/or treating fever with thrombocytopenia syndrome virus.
Test example 1
Cytotoxicity assays for Nifedipine
In Vero cells, Nifedipine was tested for cytotoxicity.
Vero cells were grown at 1X 104Cells/well were seeded in 96-well plate cell culture plates at 37 ℃ with 5% CO2Incubate for 12-16h in an incubator, treat with 50. mu. Mol/L, 100. mu. Mol/L, 200. mu. Mol/L, 300. mu. Mol/L, and 400. mu. Mol/L of Nifedipine (all DMSO as solvents), three replicates per set, and add the same amount of dimethyl sulfoxide (DMSO) to the control. After 24h of drug action, the cells were analyzed for cell viability by staining with MTT, detecting OD (optical sensitivity) 492nm, and the results are shown in FIG. 1. In FIG. 1, CC of Nifedipine50(concentration required to cause half-cell toxicity) is greater than 250. mu. Mol/L. As can be seen from FIG. 1, Nifedipine has low cytotoxicity and good biological safety.
Test example 2
Effect of Nifedipine on SFTSV replication
Vero cells were grown at 2X 105Cells/well were seeded in 24-well plates at 37 ℃ with 5% CO2Incubate for 12-16h in an incubator, treat with Nifedipine at concentrations of 25, 50, 75, 100 and 150 μ M, and add the same volume of DMSO as the negative control. After incubation for 1h, SFTSV virus infection is respectively carried out, after 1h at 37 ℃, a DMEM culture medium with a culture medium of 2% FBS is replaced, after 16h, the effect of Nifedipine on inhibiting virus replication is detected by a real-time fluorescence quantitative PCR (qPCR) method, the result is shown in figure 2, and the compound Nifedipine has a remarkable inhibiting effect on SFTSV replication as can be seen from figure 2. Under the condition of 150 mu Mol/L concentration, the compound Nifedipine has obvious inhibition effect on the replication of SFTSV.
Test example 3
Detection of SFTSV antiviral activity by Nifedipine
Vero cells were grown at 2X 105Cells/well were seeded in 24-well plates at 37 ℃ with 5% CO2Culturing in incubator for 12-16h with use concentration of 1Mu. Mol/L, 5. mu. Mol/L, 10. mu. Mol/L, 20. mu. Mol/L and 40. mu. Mol/L of Nifedipine, and the same volume of dimethyl sulfoxide (DMSO) was added as a negative control. After 10h of incubation, SFTSV is used for infection, an incubator is incubated at 37 ℃ for 1h, a DMEM (Dulbecco's modified Eagle's medium) culture medium with a culture medium of 2% FBS is replaced, after 16h, the effect of Nifedipine on inhibiting virus replication is detected through titer, and as shown in figure 3, the compound Nifedipine has a remarkable inhibiting effect on SFTSV replication under different concentration conditions. Further, IC50(half inhibition concentration) of Nifedipine inhibiting SFTSV was examined. The results of the detection are shown in FIG. 3.
From the inhibition curves of SFTSV in fig. 3, Nifedipine significantly inhibited the activity of SFTSV, and its IC50 was calculated to be 98 μ M.
Test example 4
Effect of Nifedipine on SFTSV infection of humanized mice
cg-Prkdcem1IDMOIl2rgem2IDMO mice (NOD-Prkdcnull IL2R γ null,
) Was used for humanized mouse construction, all were raised in an SPF (Specific Pathologen Free, SPF) environment, and 2 days prior to graft modification, mice were treated with intraperitoneal injection of busulfan (20 mg/kg). Subsequently, CD34 purified from human cord blood with purity of more than 90%+The cells were transplanted into mice by tail vein injection to obtain a humanized mouse model.
The humanized mice were divided into 2 groups: (1) vehicle control group: SFTSV + vehicle group (number of mice 7), (2) experimental group: SFTSV + Nifedipine (number of mice 6).
Two groups of mice are infected by an intraperitoneal injection method, and the dose of the virus of the SFTSV is 200 mu L. The experimental group was administered by gavage with a dose of Nifedipine of 100mg/kg/d twice daily for a total of 10 days. The menstruum control group is a DMEM medium with the same volume as the perfusate.
The overall experimental procedure followed strictly the guidelines of the national institutes of health of the united states as set forth in the guidelines for the operation of the animal care and use committee. The mouse daily monitoring indexes include: arch, back, hair, activity, stress, weight, body temperature, etc. Mortality was recorded in both groups of mice and the results are shown in figure 4.
As shown in fig. 4, the mortality rate of mice in the administration group was significantly reduced to 0% compared to 57.1% in the control group. The Nifedipine can obviously inhibit the replication of SFTSV at the mouse level, so that the survival rate of infected mice is obviously improved.
Test example 5
Effect of Nifedipine on SFTSV infected patients
In 2011-2017 years, the 154 hospital of the people liberation military in China receives 2087 total patients with SFTSV infection, and the clinical curative effect of nifedipine in treating SFTSV infection is analyzed by retrospective analysis research design. In a retrospective analysis, 136 patients involved in other clinical trials were first eliminated, and 135 failed to provide Ca2+The patients with channel inhibitor drug use cases secondarily exclude 12 cases of using other Ca2+The patients who had been treated with Nifedipine before the trial was performed, and the patients who had been treated with Nifedipine during the trial were selected as 83 patients (Nifedipine-treated group) and 48 patients (non-Nifedipine treated group) who had been treated with Nifedipine before the trial was performed, but had no Nifedipine during the trial), and the inclination matching method (3: 1 matching was performed according to the age, sex, delay time from onset to hospitalization) was used from 1673 patients who had never used Ca2+Of the patients with channel inhibitor drugs, 249 cases were randomly selected as general drug groups (general SFTS group, before and after admission, without any Ca use)2+Channel inhibitor class of drugs for treatment). The institutional ethics committee approval is obtained before the research is carried out, the national medical research standard is strictly observed all the time, and the written consent of the patient is required in advance by all sampling related operations.
Continuous variable comparisons between groups were performed using a t-test or a non-parametric test (Mann-Whitney test), and categorical variable comparisons between groups were performed using a chi-square test or Fisher's exact test. Comparative analysis of baseline data found: the difference in age, sex, and delay time from onset to hospitalization of the three groups of patients was not statistically significant (all P > 0.05); the difference in SFTSV viral load at admission was not statistically significant for the three groups of patients (all P > 0.05); the difference in supportive treatment received during hospitalization of the three groups of patients was not statistically significant (all P > 0.05). Survival analysis was performed by Kaplan-Meier method (see FIG. 5), and the mortality rate of the Nifedipine treatment group was found to be 3.6%, which was significantly lower than that of the general drug group (mortality rate 19.7%, P <0.001) and the non-Nifedipine treatment group (mortality rate 20.8%, P < 0.001).
From test examples 1 to 5, it is clear that Nifedipine (Nifedipine) inhibits the replication of SFTSV. SFTSV infected mice were significantly reduced in mortality and weight change following Nifedipine administration. The curative rate of clinical SFTSV infected patients can be remarkably improved by Nifedipine.
In addition, nifedipine shows definite antiviral effect in cell, animal level and clinic, has good application prospect, and can be used for preparing anti-SFTSV drugs.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (9)
1. Use of a composition comprising nifedipine or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention and/or treatment of fever with thrombocytopenia syndrome virus.
2. The use according to claim 1, wherein the composition comprises from 50 μ Mol/L to 400 μ Mol/L of said nifedipine or a pharmaceutically acceptable salt thereof.
3. The use according to claim 1, wherein the composition comprises 120 μ Mol/L to 170 μ Mol/L of the nifedipine or the pharmaceutically acceptable salt thereof.
4. Use according to claim 1, wherein said composition comprises from 150 μ Mol/L to 155 μ Mol/L of said nifedipine.
5. The use according to claim 1, wherein the composition is for the manufacture of a medicament for inhibiting replication of the fever with thrombocytopenia syndrome virus.
6. The use of claim 1, wherein the composition further comprises a pharmaceutically acceptable adjuvant.
7. The use according to claim 1, wherein the composition is in the form of a tablet.
8. The use of claim 1, wherein the medicament is in a dosage form for gastrointestinal administration.
9. The use of claim 1, wherein the medicament is administered orally.
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