CN108042532B

CN108042532B - Antifungal pharmaceutical composition

Info

Publication number: CN108042532B
Application number: CN201711116808.6A
Authority: CN
Inventors: 任彪; 朱乘光; 程磊; 周学东
Original assignee: Sichuan University
Current assignee: Sichuan University
Priority date: 2017-11-13
Filing date: 2017-11-13
Publication date: 2021-05-18
Anticipated expiration: 2037-11-13
Also published as: CN108042532A

Abstract

The invention discloses an antifungal medicine composition, which comprises an antifungal A medicine and a synergistic B medicine; the fungus inhibition concentration coefficient FICI of the medicine composition is equal to (MIC of A medicine in the medicine composition/MIC of A medicine when the A medicine is used alone) + (MIC of B medicine in the medicine composition/MIC of B medicine when the B medicine is used alone) and is less than or equal to 0.5, wherein the A medicine comprises econazole, and the B medicine comprises artemisinin. The invention can effectively reduce the Minimum Inhibitory Concentration (MIC) of the antifungal drug econazole and can obviously improve the antifungal effect of econazole, thereby reducing the dosage of econazole in the treatment process and further reducing the toxic and side effects of econazole.

Description

Antifungal pharmaceutical composition

Technical Field

The invention belongs to the technical field of antifungal treatment, and particularly relates to an antifungal pharmaceutical composition.

Background

Fungal infections include superficial fungal infections with high incidence but low risk and deep fungal infections with low incidence but high risk. The treatment of fungal infections is faced with serious challenges due to the increasing incidence of antibiotic abuse, especially opportunistic fungal infections in immunocompromised patients, in recent years. Although new antifungal medicines are continuously available and provide new opportunities for antifungal treatment, the reports of drug resistance related to drug treatment are gradually increased. There is therefore a need to develop new antifungal therapies.

Pyrrole antifungal drugs, also known as azole antifungal drugs, are the antifungal drugs currently most widely used clinically, and are also a class of antifungal drugs with better prospects, and are mainly classified into imidazoles and triazoles by inhibiting the biosynthesis of sterols in fungal cell membranes. Econazole is a representative drug of azole antibiotics. The pyrrole antifungal medicine has the advantages of good curative effect, wide antibacterial spectrum, less toxicity and the like compared with amphotericin B. However, in recent years, the curative effect of the pyrrole antifungal drugs is reduced and even the drug resistance phenomenon is caused due to the abuse phenomenon of antibiotics and antifungal drugs, meanwhile, the pyrrole antifungal drugs have certain hepatotoxicity and hepatotoxicity, in recent years, a plurality of cases of hepatotoxicity and even death are reported, and some pyrrole antifungal drugs have adverse reactions such as gastrointestinal toxicity, genital toxicity, teratogenic mutation, abnormal vision, rash, fever, headache, hallucination, nausea, vomiting, abdominal pain and the like. Therefore, the research and development of the high-efficiency and low-toxicity pyrrole antifungal medicine composition is the key point of the development of antifungal medicines in a period of time in the future.

Disclosure of Invention

The invention aims to provide an antifungal medicine composition, and aims to solve the problems of lack of antifungal medicine composition and large toxic and side effects of antifungal medicines in the prior art.

The invention is realized by an antifungal medicine composition, which comprises an antifungal A medicine and a synergistic B medicine; wherein, the fungus inhibition concentration coefficient FICI of the medicine composition is not more than 0.5 (MIC of A medicine in the medicine composition/MIC of A medicine when the A medicine is used alone) + (MIC of B medicine in the medicine composition/MIC of B medicine when the B medicine is used alone).

Preferably, the a drug comprises a pyrrole antibiotic, or an analogue, derivative, prodrug, metabolite and pharmaceutically active salt of a pyrrole antibiotic.

Preferably, the B drug comprises a sesquiterpene lactone drug, or analogs, derivatives, prodrugs, metabolites and pharmaceutically active salts of a sesquiterpene lactone drug.

Preferably, the A drug is econazole and the B drug is artemisinin.

Preferably, the mass ratio of the econazole to the artemisinin is (0.125-0.5): (1.5625-100).

Preferably, the mass ratio of the econazole to the artemisinin is 0.125: 100.

preferably, the fungi include superficial fungi that are less harmful to the body and deep fungi that are more harmful.

Preferably, the fungus comprises candida albicans.

Compared with the defects and shortcomings of the prior art, the invention has the following beneficial effects: the invention can effectively reduce the Minimum Inhibitory Concentration (MIC) of the antifungal drug econazole and can obviously improve the antifungal effect of econazole, thereby reducing the dosage of econazole in the treatment process and further reducing the toxic and side effects of econazole.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

It should be understood that the experimental methods used in the following examples are all conventional methods unless otherwise specified, and that the materials, reagents, etc. used in the following examples are commercially available without otherwise specified.

Candida albicans SC5314 was purchased from American Type Culture Collection (American Type Culture Collection) and deposited as ATCC MYA-2876.

Artemisinin was purchased from carbofuran corporation (catalog No. 949289-1 g).

Econazole is available from MCE corporation (Cat. No. HY-B0885).

Interactive antifungal Activity test for artemisinin and econazole

The interactive antifungal (Candida albicans sc5314) activity of artemisinin and econazole was tested using the checkerboard method.

1. Candida albicans SC5314 is cultured in RPMI1640 medium at 35 deg.C and 80% humidity, and 5% CO₂Under the conditions of (1).

2. Dissolving artemisinin and econazole in DMSO respectively to concentrations of 20mg/ml and 8mg/ml respectively, and storing in refrigerator for use.

3. Determination of Minimum Inhibitory Concentration (MIC)

(1) Reference is made to the protocol of the Clinical and Laboratory Standardization Institute (CLSI) M27-A3, the "Yeast liquid Medium dilution antifungal drug sensitivity test protocol". A series of diluted artemisinin and econazole solutions were prepared from the stock 20mg/ml artemisinin and 8mg/ml econazole solutions in liquid medium (RPMI 1640 medium).

(2) Extracting Candida albicansCells were seeded in 96-well plates at 96ul per well (about 1X 10)⁴Individual cells), 2ul artemisinin and econazole liquid medicine were added, respectively, the combination method of the liquid medicine was as follows:

the first drug econazole is longitudinally arranged on a 96-well plate from top to bottom according to the dilution concentration (the econazole concentration of each horizontal row is the same), each hole is 2ul, the second drug artemisinin is transversely arranged from left to right according to the dilution concentration (the artemisinin concentration of each vertical row is the same), each hole is 2ul, the concentration of each empty artemisinin and econazole is recorded, a group which is independently added with 2ul of artemisinin or econazole liquid medicine with gradient dilution and a blank control group which is not added with any medicine are simultaneously arranged, three parallel 96-well plates are made, and the results are averaged.

(3) And (3) incubating the 96-well plate for 16-20 hours at 35 ℃, and then testing the OD value at 600 nm.

(4) The MIC value of the azole antifungal drug is defined as the lowest drug concentration capable of obviously inhibiting the growth of fungi. The MIC values were determined for each drug combination well by comparison to the OD values of the blank.

The following definitions are provided here: whether the two drugs A and B act synergistically, additively or antagonistically is judged by inhibiting the concentration coefficient FICI.

Wherein, FICI (A/MICA alone in MIC drug combination) + (B/MICB alone in MIC drug combination), if the FICI value is less than or equal to 0.5, the synergistic effect exists between the drugs A and B, if the FICI value is between 0.5 and 4.0, the additive effect of the activities of the drugs A and B is shown, and if the FICI value is more than 4.0, the antagonistic effect exists between the drugs A and B.

The interaction of artemisinin and econazole was tested by the above definition and due to the poor anti-candida albicans activity of artemisinin itself, it (artemisinin/MIC artemisinin in the MIC drug combination) was negligible and the results are shown in table 1.

TABLE 1 FICI after combination of artemisinin and econazole

Table 1 shows that the final concentration of econazole is 0.500ug/ml, and the final concentration of artemisinin is 1.5625 ug/ml; the final concentration of econazole is 0.500ug/ml, and the final concentration of artemisinin is 3.125 ug/ml; the final concentration of econazole is 0.500ug/ml, and the final concentration of artemisinin is 6.25 ug/ml; the final concentration of econazole is 0.250ug/ml, and the final concentration of artemisinin is 12.5 ug/ml; the final concentration of econazole is 0.250ug/ml, and the final concentration of artemisinin is 25 ug/ml; the final concentration of econazole is 0.125ug/ml, and the final concentration of artemisinin is 50 ug/ml; in the seven groups of medicine combination with the final concentration of the econazole being 0.125ug/ml and the final concentration of the artemisinin being 100ug/ml, the econazole and the artemisinin have the synergistic antifungal effect. Wherein the synergistic antifungal activity of econazole and artemisinin is increased with increasing final concentration of artemisinin.

Interactive antifungal of di-and pyrrole antibiotics and semi-terpene lactones

Econazole may be replaced with azole antibiotics or analogs, derivatives, prodrugs, metabolites and pharmaceutically active salts thereof. Artemisinin can be replaced by sesquiterpene lactones and analogs, derivatives, prodrugs, metabolites and pharmaceutically active salts thereof.

The selection of more specific components between the pyrrole antibiotics and the semi-terpene lactones is selected and determined according to the type of fungi to be inhibited, the fungi which can be inhibited by the pharmaceutical composition provided by the invention are wide in source, including but not limited to superficial fungi which are slightly harmful to organisms and deep fungi which are seriously harmful, and the optimal inhibition effect of the pharmaceutical composition on the fungi can be obtained by determining the optimal mixing ratio of the two through the implementation method.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.

Claims

1. An antifungal drug composition is characterized in that the drug composition consists of an antifungal drug A and a synergistic drug B; wherein, the fungus inhibitory concentration coefficient FICI of the medicine composition is (the minimum inhibitory concentration of the A medicine in the medicine composition/the minimum inhibitory concentration of the A medicine when the A medicine is used alone) + (the minimum inhibitory concentration of the B medicine in the medicine composition/the minimum inhibitory concentration of the B medicine when the B medicine is used alone), and the FICI is less than or equal to 0.5;

the A medicine is econazole, and the B medicine is artemisinin;

the mass ratio of the econazole to the artemisinin is (0.125-0.5): (1.5625-100);

the fungus is Candida albicans.

2. The antifungal pharmaceutical composition of claim 1 wherein the mass ratio of econazole to artemisinin is 0.125: 100.