CN110856722A

CN110856722A - Use of cyclic target amine or derivative thereof

Info

Publication number: CN110856722A
Application number: CN201810977013.2A
Authority: CN
Inventors: 苑纯秀; 许静秀; 冯新港; 张佳男; 贾英英
Original assignee: Shanghai Veteromaru Research Institute Caas China Animal Health And Epidemiology Center Shanghan Branch Center
Current assignee: Shanghai Veteromaru Research Institute Caas China Animal Health And Epidemiology Center Shanghan Branch Center; Shanghai Veterinary Research Institute CAAS
Priority date: 2018-08-26
Filing date: 2018-08-26
Publication date: 2020-03-03

Abstract

The invention discloses an application of cyclotargmine or a derivative thereof in preparing a medicament for preventing or treating schistosomiasis. The invention also discloses a pharmaceutical composition containing the cyclotargmine or the derivative thereof. The cyclotardamine or the derivative thereof can kill the schistosoma japonicum schistosomulum for 14 days at the concentration of more than 1 mu M, has stronger function of killing the schistosoma japonicum schistosomulum, is expected to become a new anti-schistosomiasis medicine, and has very wide application prospect.

Description

Use of cyclic target amine or derivative thereof

Technical Field

The invention relates to the technical field of biological medicines, in particular to a new application of cyclotargmine or a derivative thereof.

Background

Schistosomiasis is parasitic disease of both human and animals caused by schistosomiasis infection, which causes serious pathological damage to multiple organs of a host, causes anemia, malnutrition, growth and development retardation, and reduced working capacity and even death. At present, schistosomiasis is reported in 78 countries and regions worldwide, about 20 million people die each year, and 8 hundred million people are at risk of contracting schistosomiasis, mainly school-age children. According to the data updated by the world health organization in the year 2017 in the october, at least 2 hundred million and 650 ten thousand people need preventive treatment in 2016, and the striking and alarming numbers warn us that schistosomiasis prevention and control work can never be relaxed.

Schistosomiasis is prevalent mainly in tropical and subtropical regions, especially in poor regions where safe drinking water is lacking and sanitary conditions are poor, with 92% of schistosomiasis patients requiring treatment living in africa. Infectious cercaria in the life cycle of schistosome live lives in fresh water, so that people mainly infected are those engaged in agriculture and fishery. However, with the migration of population and the change of environment, the epidemic range of schistosomiasis can be expanded. Schistosomiasis suffers from both human and animal suffering, and many host insects are protected, wherein more than 40 kinds of mammals can naturally infect the schistosoma japonicum. These animal sources of infection are difficult to manage and control, increasing the difficulty of preventing and controlling schistosomiasis. The control and elimination of schistosomiasis are still far from the control situation of the global schistosomiasis at present.

Over the last 30 years, the only chemotherapeutic drug praziquantel has been relied upon globally for the prevention and control of schistosomiasis. Praziquantel plays an important role in the prevention and control of global schistosomiasis, but praziquantel also has some limitations. The main performance is as follows: (1) praziquantel has weak or ineffective killing power to the juvenile insect, and the juvenile insect can still survive and generate pathogenicity in the presence of the drug. Large-scale treatment can realize 60-90% reduction of the load number of the schistosome, but cannot achieve the effect of completely eliminating the schistosome. (2) Praziquantel belongs to the racemate, and half of the dose used is pharmacologically ineffective. (3) The praziquantel absorbed into the body can be quickly metabolized into an ineffective metabolite through hydroxylation, only limited medicines can enter blood circulation to directly contact with the schistosome, the medicine action time is short, and the insecticidal action is not lasting. In addition, there is a risk of drug resistance arising from the repeated use of praziquantel in large quantities over an extended period of time. Partial severe epidemic areas of Schistosoma mansoni have successively reported that the efficacy of praziquantel is reduced, and a Schistosoma mansoni praziquantel resistant field strain is separated in patients in the epidemic areas. Further research finds that the resistance character of the schistosoma mansoni praziquantel is dominant heredity, which means that the resistance to the praziquantel can spread along with the re-propagation of schistosoma. Although the praziquantel resistant strain is not separated in the schistosome epidemic area in China in the present field, the schistosoma japonicum praziquantel resistant strain is induced by repeated sub-dose treatment in a laboratory, which shows that the schistosoma japonicum can also generate drug resistance under the continuous drug pressure of the praziquantel. And compared research of the resistant strain and the sensitive strain shows that the pathogenic and transmission capability of the resistant strain is stronger. These findings are increasing concern about schistosomiasis resistance, and the development of a wide range of resistance would pose a great threat to global schistosomiasis control without other safe and effective alternatives. The urgent need is to actively develop new anti-schistosome drugs, screen drugs which can effectively kill schistosomulum and have lasting drug effect, and make up the limitation of praziquantel action. The fluke inhibitor is combined with praziquantel or used alternatively to achieve the effect of completely eliminating schistosome in a host body.

Cyclobenzamine (Cyclopamine) is a natural steroid alkaloid extracted from veratrum plants, and is found in the mid-nineteenth century, the united states department of agriculture, when studying facial deformities of the cranium of sheep frequently occurring in the southwestern region of edhol. After genetic factors are eliminated, the fact that the ewes ingest the caladium veratrum containing the cyclotargmine 8-17 days after pregnancy is finally determined to be the direct cause of the congenital malformation of the lambs, and the period is just the stage of the formation of the nerve tube and the formation of the embryo pattern in the process of sheep embryo development. And the ewes or the rams in the time periods other than 8-17 days of pregnancy can not generate craniofacial malformed lambs when eating the California veratrum, and can not generate toxicity to the ewes and the rams. The ring targets were successfully isolated and formally named in 1968 by veratrum californica. After thirty years, Beachy et al discovered that Shh gene knock-out mice produced craniofacial malformation phenotypes similar to that of cyclotargetine when studying the regulatory function of the Hedgehog signaling pathway, and speculated that cyclotargetine may have inhibitory effects on the Hh pathway. Chen et al, 2002, reported the use of fluorescence binding experiments to confirm that cyclic-statins block the transmission of the Hh pathway by binding directly to its Smo receptor. The cyclic target is further applied to the functional research of the Hh pathway as an inhibitor of the Hh signaling pathway. The liver cancer is related to the abnormal high expression of the Hh pathway members, and researches show that the cyclotaraxane can reduce the survival capability of liver cancer cells by inhibiting the transmission of the Hh signal pathway and inducing apoptosis. Similarly, treatment with cyclotaraxane also induces apoptosis in colorectal cancer tumor cells.

The previous research on the target has focused on the tumor inhibition effect of the drug, and no published report on the use of the target as an anti-schistosome drug has been found so far.

Disclosure of Invention

The invention aims to solve the technical problem of lack of new anti-schistosomiasis drugs at present and provides a new application of cyclotargmine or a derivative thereof in preparing drugs for preventing and treating schistosomiasis.

In order to solve the technical problems, the invention is realized by the following technical scheme:

in one aspect of the invention, the invention provides an application of cyclotargmine or a derivative thereof in preparing a medicament for preventing or treating schistosomiasis.

In another aspect of the invention, an anti-schistosome drug is also provided, which comprises cyclotargmine or a derivative thereof.

In another aspect of the present invention, there is also provided an anti-schistosomiasis pharmaceutical composition, comprising a safe and effective amount of cyclotardamine or its derivative, and a pharmaceutically acceptable carrier.

The acceptable carrier is non-toxic, can be administered adjunctively and does not adversely affect the therapeutic efficacy of the cyclic target amine or derivative thereof. Such carriers can be any solid excipient, liquid excipient, semi-solid excipient, or in aerosol compositions, gaseous excipient, commonly available to those skilled in the art.

The cyclotargmine or derivative thereof of the present invention is administered in a therapeutically effective amount by oral, systemic (e.g., transdermal, nasal inhalation, or by suppository), or parenteral (e.g., intramuscular, intravenous, or subcutaneous). The preferred mode of administration is oral, which may be modulated by the extent of the disease.

Various dosage forms of the pharmaceutical composition of the present invention can be prepared according to conventional methods in the pharmaceutical field. For example, the cyclotargmine (active ingredient) is mixed with one or more carriers and then formulated into the desired dosage form, e.g., tablets, semi-solids, powders, solutions, suspensions, formulations, and the like.

In another aspect of the invention, the application of the cyclic target amine or the derivative thereof and the praziquantel in preparing the medicament for preventing or treating schistosomiasis is also provided.

The cyclic target amine or the derivative thereof can be combined with praziquantel to be used for preventing or treating schistosomiasis.

In another aspect of the invention, the invention also provides an anti-schistosome compound preparation, which comprises cyclotaramine or derivatives thereof and praziquantel. The cyclotargmine or the derivative thereof can effectively kill schistosomulum and just make up the limitation that praziquantel has weak or ineffective killing power on schistosomulum, so the compound medicine containing the cyclotargmine and the praziquantel can more effectively kill schistosomes and prevent and control schistosomiasis.

In the present invention, the term "ring-targeted derivative" refers to a compound derived by substituting a hydrogen atom or an atomic group in a ring-targeted compound with another atom or group, which has the same or similar activity.

The cyclotardamine or the derivative thereof can kill the schistosoma japonicum schistosomulum for 14 days at the concentration of more than 1 mu M, has stronger function of killing the schistosoma japonicum schistosomulum, is expected to become a new anti-schistosomiasis medicine, and has very wide application prospect.

Drawings

The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.

FIG. 1 is a diagram showing a comparison of the state of Schistosoma japonicum cultured in vitro with 10. mu.M of the loop target in example 1 of the present invention for 24 hours and 14 days;

FIG. 2 is a diagram showing a comparison of the state of Schistosoma japonicum cultured in vitro with 5. mu.M of the loop target in example 1 of the present invention for 24 to 60 hours and 14 days;

FIG. 3 is a diagram showing a comparison of the state of 1. mu.M of the loop target of example 1 of the present invention cultured in vitro for 14 days and 24 hours in the presence of Schistosoma japonicum.

Detailed Description

Example 1 anti-schistosome experiment of cyclic Tarming

1. Material method

1.1 Collection of Schistosoma japonicum materials

The Anhui cercaria ventral patch is infected with healthy New Zealand white rabbit, and the patch is kept for 20 minutes, during which time some water should be properly added to the patch part for moistening. The number of infections per rabbit was approximately 5000. The killing was performed 14 days after infection. Cercaria parasitize in hepatic portal vein and mesenteric vein, flush out larva by hepatic portal vein perfusion, lightly flush larva body with PBS (phosphate buffer solution) sterilized under high pressure and added with anticoagulant, and remove impurities and blood cells mixed in larva.

1.2 Range determination of effective concentration of Cyclic Tarming

1.23mg of cyclotaraxminstrobin powder was dissolved in 150. mu.l of ethanol to a final concentration of 20mM and stored at-20 ℃. An appropriate amount of 14-day schistosomulum is placed in a 12-hole culture plate, and 2ml of 1640 culture medium containing 10% FBS is added into each hole for culture. 37 ℃ and 5% CO₂Under the culture condition, after the polypide is cultured for 24h, cyclotetramine (20mmol/L) is added into the culture medium, so that the final concentration is 1 mu mol/L, 5 mu mol/L and 10 mu mol/L respectively. Corresponding ethanol solvent control groups are arranged, and 3 groups are respectively arranged between the experimental group and the control group. The culture medium is changed regularly, target-binding enzyme with corresponding concentration is added, the activity and the survival state of the larva are observed regularly, and the relationship between the concentration of the target-binding enzyme and the killing capacity of the larva to blood fluke is established.

2. Results

2.1 As shown in FIG. 1, 10. mu.M of the ring-target was able to cause all the 14-day juvenile worms cultured in vitro to die within 24 hours, and under the microscope, black crystals were seen to penetrate from the inside of the worm body. The activity of the insect bodies of the ethanol solvent control group with the same concentration is not abnormal.

2.2 As shown in figure 2, the 14-day juvenile worms cultured in vitro with 5 μ M cyclotaraxminshen violaceum were apparently stiff under a microscope after 24 hours, and black lumps scattered in the worm were present; after 36 hours, the content in the intestinal canal of the polypide is obviously solidified, and does not flow back and forth in the intestinal canal along with the bending and extending of the polypide like a normal polypide, and the sucker of the polypide shrinks and cannot be attached to the culture plate; after 48 hours, the worm body is more rigid and can hardly bend and stretch; the pests died and blackened after 60 hours. After the addition of the cyclotardamine is stopped, the bodies cannot be recovered to be normal, and all the bodies die within 5 days. The activity of the insect bodies of the ethanol solvent control group with the same concentration is not abnormal.

2.3 As shown in figure 3, 1 μ M of cyclotargmine causes the internal solidification of the worm intestine within 24 hours, the worm is rigid and can not freely bend and stretch, the solidification in the worm intestine is intensified along with the extension of the culture time, the sucking disc gradually shrinks and loses the adsorption capacity, the worm can not recover to the normal state after the addition of the cyclotargmine is stopped, and all the worms die within 5 days. The growth state of the polypide in the control group cultured in parallel is good.

3. Conclusion

The target-binding agent with concentration of more than 1 μ M can kill schistosoma japonicum schistosomulum within 5 days in vitro culture, has effect in killing schistosoma japonicum schistosomulum, and can be used as candidate drug of new anti-schistosomiasis medicine.

The above-mentioned embodiments only express the embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. The application of the cyclic target amine or the derivative thereof in preparing the medicine for preventing or treating schistosomiasis.

2. An anti-schistosome medicine contains cyclotargmine or its derivative.

3. An anti-schistosome pharmaceutical composition comprises a safe and effective amount of cyclotargmine or derivatives thereof and a pharmaceutically acceptable carrier.

4. Use of the pharmaceutical composition of claim 3 for the preparation of a medicament for the prevention or treatment of schistosomiasis.

5. The application of the combination of the cyclic target amine or the derivative thereof and the praziquantel in preparing the medicine for preventing or treating schistosomiasis.

6. A compound preparation for resisting schistosomiasis contains cyclotardamine or its derivative and praziquantel.