阅读说明：本技术 含有吡咯亚基二氢吲哚酮衍生物混合物的组合物及其用途 (Compositions containing mixtures of pyrrolylideneindolinone derivatives and uses thereof ) 是由 向飞 李海燕 于 2019-11-19 设计创作，主要内容包括：本发明提供了一种含有互不相同的第一吡咯亚基二氢吲哚酮衍生物与第二吡咯亚基二氢吲哚酮衍生物的混合物的组合物,所述的混合物能与穿破石、毛老虎、六月雪、草苁蓉、红药子、定心荣、山八角等植物提取物对产碱性纤维素酶浅黄金色单胞菌等病原菌产生协同的抗菌作用。(The invention provides a composition containing a mixture of a first pyrrolizidine indolinone derivative and a second pyrrolizidine indolinone derivative which are different from each other, wherein the mixture can generate a synergistic antibacterial effect with plant extracts such as cudrania cochinchinensis, cutworm, serissa serissoides, cistanche salsa, rhodochrous japonicas, xylocarpus parviflorus, illicium simonsii and the like on pathogenic bacteria such as golden yellow monad producing alkaline cellulase.)

1. A composition comprising a mixture of a first pyrrolizidine indolinone derivative and a second pyrrolizidine indolinone derivative which are different from each other and selected from the group consisting of compounds 1 to 12 shown below:

2. the composition according to claim 1, wherein the mass ratio of the first pyrrolizidine indolinone derivative to the second pyrrolizidine indolinone derivative is 0.01:1 to 100: 1.

3. The composition of claim 1 or 2, wherein said composition further comprises a plant extract.

4. The composition of claim 3, wherein said plant extract is an extract of a plant selected from the group consisting of Cudrania cochinchinensis, Tiger, Serissa japonica, cistanche salsa, Rad mercuric oxide, and Illicium verum.

5. The composition of claim 1 or 2, wherein said composition is formulated as an oral solid dosage form.

6. The composition of claim 5, wherein said oral solid dosage form is one selected from the group consisting of a tablet, a capsule and a capsule.

7. Use of a composition according to claim 1 or 2 for the preparation of a medicament for the treatment of bacterial infectious diseases.

8. Use according to claim 7, characterized in that the bacterial infectious disease is a disease caused by an infection with a bacterium selected from the group consisting of alkaline-producing cellulase aureomonas euryales, acinetobacter junii, clostridium perfringens, staphylococcus intermedia, staphylococcus cohnii, staphylococcus haemolyticus, lissakei, streptococcus oralis, listeria monocytogenes, pantoea agglomerans, acinetobacter lofoenii, corynebacterium mimicus, moraxella catarrhalis, alcaligenes xylooxidans, staphylococcus pasteurii, streptococcus anaerobically digested, morganella morganii, proteus mirabilis, streptococcus magnus, legionella pneumophila.

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a composition containing a mixture of pyrrylidene indolinone derivatives and application thereof.

Background

Nosocomial infections are a significant problem affecting hospital medical quality. With the continuous development and progress of modern medical technology, a large amount of antibacterial drugs are used, and the drug-resistant strains of clinical pathogenic bacteria are continuously increased. Therefore, the improvement of the antibacterial activity of the existing drugs can help to improve the treatment effect of the patients with bacterial infection.

CN108939081A discloses various protein tyrosine kinase inhibitors with antibacterial activity, including pyrrolylideneindolinone derivatives represented by formula i.

The plant extract is rich in a plurality of components with antibacterial activity, for example, the extracts of plants such as the populus nymphs and the like (the report of tropical biology, 2019,10(03): 226;) 230. the extracts of the plants such as the blumea riparia, the rhinestone, the illicium verum, the aeolian milkvetch root, the sinoacutus hexapetalus mongholicus, the kadsura longipedunculata, the anoectochilus roxburghii, the typhonium giganteum, the rhynchophyllum giganteum, the stringy euphorbia root, the typhonium giganteum, the fraxinellus pallidus, the cudrania cochinchinensis, and the like have certain antibacterial activity.

It is well known that the therapeutic effects of Chinese herbs are manifested by the synergistic action of a series of compounds (effective components) with similar structures. Therefore, a compound group which can generate synergistic action with the plant extract is searched, and a new way is provided for treating difficult and complicated diseases by combining Chinese and western medicine.

Disclosure of Invention

The invention aims to provide a composition containing a mixture of a first pyrrolizidine indolinone derivative and a second pyrrolizidine indolinone derivative which are different from each other, wherein the mixture can generate a synergistic antibacterial effect with extracts of plants such as cudrania cochinchinensis, boston ivy, serissa serissoides, cynanchum otophyllum, gordonia macrogola, illicium dunnianum and the like.

In order to achieve the above objects, the present invention provides, in one aspect, a composition comprising a mixture of a first pyrrolizidine indolinone derivative and a second pyrrolizidine indolinone derivative, which are different from each other and selected from the following compounds 1 to 12:

in one aspect, the mass ratio of the first pyrrolizidine indolinone derivative to the second pyrrolizidine indolinone derivative in the composition of the invention is preferably 0.01: 1-100: 1.

In another aspect, the composition of the present invention preferably further comprises a plant extract.

More preferably, the plant extract of the present invention is an extract of one plant selected from the group consisting of cudrania cochinchinensis, boston ivy, serissa serissoides, boston stephania root, naberry, and illicium dunnianum.

In another aspect, the composition of the present invention can be prepared into oral solid preparation.

More preferably, the oral solid preparation of the present invention is one selected from the group consisting of tablets, capsules and capsules.

In another aspect, the present invention provides the use of a composition as described above for the preparation of a medicament for the treatment of a bacterial infectious disease.

Preferably, the bacterial infectious disease of the present invention is a disease caused by infection with one bacterium selected from the group consisting of alkaline cellulase-producing aureomonas, acinetobacter juni, clostridium perfringens, staphylococcus intermedius, staphylococcus cohnii, staphylococcus haemolyticus, otitis externa, streptococcus oralis, listeria monocytogenes, pantoea agglomerans, acinetobacter lofoenii, corynebacterium mimicus, moraxella catarrhalis, alcaligenes xylosoxidans, staphylococcus pasteurii, streptococcus anaerobicus, morganella morganii, proteus mirabilis, streptococcus magnus, legionella pneumophila.

In vitro test results show that the mixture of the pyrrolizidine indolinone derivative can generate synergistic antibacterial action with extracts of multiple plants such as cudrania cochinchinensis, cutworm, serissa serissoides, cynanchum otophyllum, gorgon europaea, illicium simonsii and the like

Detailed Description

The following description of the embodiments is only intended to aid in the understanding of the method of the invention and its core ideas. It should be noted that, for those skilled in the art, it is possible to make various improvements and modifications to the present invention without departing from the principle of the present invention, and those improvements and modifications also fall within the scope of the claims of the present invention. The following description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are now described.

Preparation example 1 preparation of plant extract

Drying the dried plant material at 40 deg.C by ultrasonic extraction, pulverizing, sieving with 40 mesh sieve, and placing into self-sealing bag. Before the test, 50g of the dried plant powder was weighed and added with different organic solvents for ultrasonic extraction (as shown in table 1), and the extraction was repeated 3 times for 60min each time. Filtering, concentrating, and storing in refrigerator at 4 deg.C.

TABLE 1 organic solvent for ultrasonic extraction of different plant extracts

Plant material	Extract number	Organic solvent
			Wind coming from corner	PE1	Methylene dichloride
Root of Teng pseudo-ginseng	PE2	Methylene dichloride
			Six-ear bell	PE3	Chloroform
Eight hundred centimetres	PE4	Methanol
			Zhui Gu Feng (wind dispelling from bone)	PE5	Methylene dichloride
All-grass of Szechwan-Yunnan Sambucus	PE6	Methanol
			Rhizoma Osmundae	PE7	Ethanol
Medicine for treating bone fracture	PE8	Ethanol
			Radix Euphorbiae Fischerianae	PE9	Ethyl acetate
All-grass of Anoectochilus roxburghii	PE10	N-heptane
			Piercing stone	PE11	Ethyl acetate
Tiger with hair	PE12	Chloroform
			Root of common June	PE13	N-heptane
Herb desert cistanche	PE14	Acetone (II)
			Hongyao Zi	PE15	Ethyl acetate
Good heart-nourishing	PE16	N-heptane
			Illicium verum	PE17	Chloroform
One-foot wind	PE18	Acetone (II)
			White diamond	PE19	N-heptane
Folium Ferri Suffruticosae	PE20	Acetone (II)

Test example 1 mixture of pyrrolylideneindolinone derivatives (Effect on antibacterial Activity of plant extracts)

The inhibition of various bacteria by the test substances ① - ③ shown below is determined by a method disclosed by Lemna hexandra et al (food and fermentation industry, 2017,43(02): 232-238.).

(1) ① test drug prepared by taking pyrrolizidine indolinone derivative compounds X and Y (X, Y is selected from 1-12, and X is not equal to Y) according to a specific mass ratio (R)₁) Mixing, namely MIX (X-Y), ②, the plant extract (PEZ, Z is 1-20) obtained in preparation example 1, ③, ① and ②, and mixing according to the mass ratio (R)₂) The resulting mixture was designated MIX (X-Y-PEZ).

(2) Test method

① preparing test sample solution by taking a certain amount of test substance, using DMF as solvent, and adopting a specific multiple continuous dilution method to prepare test sample solution with 6 concentrations.

② bacterial suspension is prepared by inoculating activated strain to liquid culture medium (no agar), shake-culturing, calculating bacterial colony number by plate dilution method, and adjusting bacterial suspension concentration to 10 with sterile physiological saline⁷CFU/mL。

③ agar-hole diffusion method for determining antibacterial activity comprises cooling the sterilized culture medium to 50 deg.C, adding 6mL of bacterial suspension, mixing, pouring into a culture dish with diameter of 9cm, standing for 46min, uniformly perforating (diameter of 7mm) on the cured culture medium with an aseptic perforator, marking, adding 40 μ L of sample solution into each hole, DMF as blank control, culturing the bacteria at 37 deg.C for 18h, measuring and recording the diameter (mm) of the zone of inhibition, repeating for 3 times, averaging to obtain the result, and calculating the Inhibition Ratio (IR) of the strain to be tested according to the following formula.

For MIX (X-Y) and PEZ, IR is plotted against the logarithm (log (c)) of the total concentration of MIX (X-Y) and the tested concentration (ng/mL) of PEZ, and the concentration of each test substance at which inhibition of a specific fa occurs is calculated from the linear regression equation and is reported as IC_fa(A)And IC_fa(PEZ). For MIX (X-Y-PEZ), the concentration of MIX (X-Y) in MIX (X-Y-PEZ) at which inhibition of a particular fa occurs is calculated according to a linear regression equation using IR plotted against the logarithm of the concentration (ng/mL) of MIX (X-Y) therein, and is designated IC_fa(mixA)。

The Combination Index (CI) at which a specific fa inhibitory rate is produced is calculated according to the following formula.

When CI <1, it means that there is synergism, the smaller the CI, the stronger the synergism.

TABLE 2.1 inhibition of the test substances on the production of alkaline cellulase aureoaureomonas sp

TABLE 2.2 test substance inhibition of Acinetobacter johnsonii

TABLE 2.3 inhibition of Clostridium perfringens by test substances

TABLE 2.4 inhibitory Effect of the test substances on Staphylococcus intermedius

TABLE 2.5 inhibitory Effect of the test substances on Staphylococcus cohnii

TABLE 2.6 inhibition of Staphylococcus hemolyticus by test substances

TABLE 2.7 test substances for their inhibitory action against the otitis media Rickettsia

TABLE 2.8 inhibitory Effect of the test substances on Streptococcus oralis

TABLE 2.9 inhibitory Effect of test substances on Listeria monocytogenes

TABLE 2.10 inhibition of Pantoea agglomerans by test substances

TABLE 2.11 inhibition of Acinetobacter lofoii by test substances

TABLE 2.12 inhibition of Corynebacterium parvum by test substances

TABLE 2.13 inhibition of Moraxella catarrhalis by test substances

TABLE 2.14 inhibitory Effect of test substances on xylose oxidizing Alcaligenes

TABLE 2.15 inhibition of Staphylococcus pasteurii by test substances

TABLE 2.16 inhibition of anaerobic digestion of Streptococcus by test substances

TABLE 2.17 inhibitory Effect of the test substances on Morganella morganii

TABLE 2.18 inhibition of Proteus mirabilis by test substances

TABLE 2.19 inhibition of Streptococcus magnus by the test substances

TABLE 2.20 inhibition of Legionella pneumophila by test substances