Application of chicoric acid in preparation of medicine for preventing or treating bad pregnancy outcome

Abstract

The invention relates to the technical field of biotechnology, in particular to application of chicoric acid (especially LCA) in preparation of a medicine for preventing, relieving and / or treating adverse pregnancy outcome caused by excessive activation of IFN-I.

Description

Technical Field

[0001] The present invention relates to the technical field of biotechnology, and in particular to the use of chicoric acid (especially LCA) in preparing medicines for preventing or treating adverse pregnancy outcomes. Background Art

[0002] Based on structural characteristics, receptors, cell sources and biological activity, interferons (IFN) are divided into three types: IFN-I, IFN-II and IFN-III. The human type I interferon family is a large family based on IFN-α, including 13 IFN-α subtypes, IFN-β, IFN-ε, IFN-κ and IFN-ω; type II interferons are called IFN-γ in humans; type III interferons include IFN-λ1, IFN-λ2, IFN-λ3 and IFN-λ4.

[0003] Prolonged and excessive activation or induction of IFN-I signaling impairs embryonic development, as observed in several autoimmune diseases, such as systemic lupus erythematosus (SLE) and Icardi-Gutierrez syndrome. IFN-I signaling is required for healthy pregnancy and plays a role in pregnancy recognition and uterine artery remodeling during pregnancy. In contrast, excessive and chronic IFN-I signaling increases the activation of IFN-induced transmembrane proteins (IFITMs), which inhibit the formation of placental syncytiotrophoblasts (STs), ultimately leading to fetal death.

[0004] Adverse pregnancy outcomes refer to all pathological pregnancy and delivery complications other than normal pregnancy, mainly including spontaneous abortion, intrauterine growth restriction, premature birth (<37 weeks), premature rupture of membranes, fetal malformation, neonatal asphyxia, low birth weight (<2500 g), admission to the neonatal intensive care unit and other adverse outcomes for pregnant women and perinatal infants.

[0005] Cichoric acid (CA), also known as dicaffeoyltartaric acid, is a natural compound with the structural formula: CA has been reported to have antioxidant and anti-inflammatory properties, with the potential to alleviate the progression of atherosclerosis. L-chicoric acid (LCA) has been reported to be a potent, selective, and reversible HIV-1 integrase inhibitor that can inhibit HIV-1 replication in tissue culture. Summary of the invention

[0006] One object of the present invention is to provide a safe, effective and precise treatment for pregnant patients with overactivated IFN-I signaling.

[0007] Preferably, a safe, effective and precise treatment is provided for pregnant SLE patients with overactivated IFN-I signaling.

[0008] Another object of the present invention is to provide chicoric acid that can prevent, alleviate and / or treat adverse pregnancy outcomes, wherein the chicoric acid is preferably LCA.

[0009] Another object of the present invention is to provide chicoric acid that can prevent, alleviate and / or treat pregnancy diseases associated with excessive activation of IFN-I signals, wherein the chicoric acid is preferably LCA.

[0010] In the embodiments of the present invention, LCA is used to verify its therapeutic or alleviating effect on adverse pregnancy outcomes in pregnant patients with overactivated IFN-I signaling, so those skilled in the art can understand that chicoric acid (CA) can also achieve the same or similar effects.

[0011] Specifically, the present invention provides the following technical solutions:

[0012] 1. Use of chicoric acid in the preparation of a medicament or kit for preventing, alleviating and / or treating adverse pregnancy outcomes.

[0013] 2. Use of LCA in the preparation of a drug or kit for preventing, alleviating and / or treating adverse pregnancy outcomes.

[0014] 3. The use according to item 1 or 2, wherein the adverse pregnancy outcome is an adverse pregnancy outcome caused by excessive activation of IFN-I.

[0015] 4. The use according to item 3, wherein the IFN-I overactivation is IFN-I overactivation caused by SLE.

[0016] 5. The use according to any one of items 1-4, wherein the adverse pregnancy outcome is an adverse pregnancy outcome caused by embryo resorption and / or developmental changes.

[0017] 6. The use according to any one of items 1-5, wherein the adverse pregnancy outcome is selected from the group consisting of spontaneous abortion, intrauterine growth restriction, developmental changes, premature birth, premature rupture of membranes, fetal malformation, neonatal asphyxia, low birth weight and neonatal intensive care unit admission.

[0018] 7. The use according to any one of items 1 to 6, characterized in that the drug or kit further comprises a pharmaceutically acceptable excipient.

[0019] 8. The use according to any one of items 1-7, wherein the medicament or kit is administered at E6.5, E9.5, E10.5 and / or E11.5, preferably at E10.5.

[0020] In the present invention, "activation" and "activation" can be used interchangeably.

[0021] In the present invention, "E+days" refers to the days of embryonic development after fertilization. The mice are mated and the vaginal plug is checked the next day. The vaginal plug is used as a sign of successful mating, and the day of successful mating is E0.5.

[0022] It is known that poly (I: C) and pristane can act as IFN-I inducers and can be used to induce autoimmune-like lesions, such as SLE.

[0023] Advantages and positive effects of the present invention:

[0024] The present invention provides the use of chicoric acid (especially LCA) in the treatment of pregnancy diseases related to excessive activation of IFN-I signals at the maternal-fetal interface (such as systemic lupus erythematosus pregnancy diseases), which has good effect and high safety. Chicoric acid (especially LCA) can be used to treat pregnancy diseases with excessive activation of IFN-I signals, including pristane-induced SLE and spontaneous SLE pregnancy diseases. Therefore, the present invention has great clinical significance for the precise treatment of pregnancy-related diseases such as SLE. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Schematic diagram of the Poly (I:C) induction model and LCA treatment.

[0026] Figure 2 A is a physical picture of the pregnancy outcome of the poly (I:C)-induced model treated with LCA; B is the statistics of adverse pregnancy outcomes in the poly (I:C)-induced model treated with LCA.

[0027] Figure 3 Schematic diagram of the pristane induction model and LCA treatment.

[0028] Figure 4 A is a physical picture of the pregnancy outcome of the pristane-induced model treated with LCA; B is the statistics of adverse pregnancy outcomes of the pristane-induced model treated with LCA.

[0029] Figure 5 A is a schematic diagram of LCA treatment of NZW mice; B is the statistics of adverse pregnancy outcomes in LCA treatment of NZW mice.

[0030] Figure 6 A shows the statistics of offspring survival rate in the poly (I:C)-induced model treated with LCA, and B shows the comparison of offspring weights in each group; C shows the statistics of offspring survival rate in the pristane-induced model treated with LCA, and D shows the comparison of offspring weights in each group. DETAILED DESCRIPTION

[0031] The following examples will help those skilled in the art to further understand the present invention, but are not intended to limit the present invention in any form.

[0032] The experimental methods in the examples, unless otherwise specified, all adopt conventional techniques in the art, and the experimental reagents are all commercially available products.

[0033] The mice used in the examples were 8-15 week old C57BL / 6J female and male mice purchased from Shanghai Slake. 8-10 week old C57BL / 6J female mice were mated with C57BL / 6J male mice, and the vaginal plug was checked the next day. The vaginal plug was used as a sign of successful mating, and the successful mating was recorded as E0.5, and "E+days" indicated the number of days of embryonic development after fertilization. NZW mice were provided by Shanghai Institute of Rheumatology, Renji Hospital Affiliated to Shanghai Jiao Tong University as a spontaneous mouse model of systemic lupus erythematosus (SLE), which can be constructed by conventional methods or can be commercially obtained.

[0034] Poly (I: C) reagent was purchased from Invivogen (Cat# tlrl-pic). Pristane was purchased from Selleck (Cat# E0198). LCA was purchased from MedChemExpress (Cat# HY-N0457A). Dimethyl sulfoxide (DSMO) was purchased from BioFroxx (Cat# 10084ML100).

[0035] Example 1. In vivo functional experiments demonstrate that LCA can treat and alleviate adverse pregnancy outcomes caused by poly (I:C)-induced IFN-I overactivation

[0036] E10.5 (10.5 days after gestation) mice were intraperitoneally injected with 7.5 mg / kg poly(I:C) to induce immune activation; 500 μl of saline was injected as a control.

[0037] E10.5 and E11.5 mice were intraperitoneally injected with 10 mg / kg LCA or the same volume of solvent (95% corn oil and 5% DMSO), and the mice were sacrificed and dissected at E12.5 to observe the status of the embryos to determine the pregnancy outcome ( Figure 1 ).

[0038] The results showed that poly (I: C) induced 44.8% of the embryos in a mouse litter to undergo embryo resorption (no surviving embryos were visible to the naked eye, instead, there were blood clots absorbed by the mother) or developmental changes (the fetal brain became significantly smaller, indicating abnormal fetal brain development; the fetal liver turned white, indicating abnormal fetal liver hematopoiesis) ( Figure 2In the experiment, only 3.2% of embryos in a litter of poly(I:C)-induced mice treated with LCA showed embryo resorption or developmental changes ( Figure 2 ). The results showed that LCA treatment could significantly alleviate the embryo resorption or developmental changes induced by poly (I:C).

[0039] On the other hand, in the poly (I:C)-induced SLE model, LCA treatment significantly increased the survival rate and body weight of offspring mice ( Figure 6 AD).

[0040] Example 2. In vivo functional experiments demonstrate that LCA can treat and alleviate adverse pregnancy outcomes caused by pristane-induced IFN-I overactivation

[0041] The SLE model was successfully established by intraperitoneal injection of 500 μl of pristane into E10.5 mice to induce IFN-I signaling activation; 500 μl of normal saline was injected as a control.

[0042] E10.5 mice were intraperitoneally injected with 10 mg / kg LCA or the same volume of solvent (95% corn oil and 5% DMSO), and the mice were sacrificed and dissected at E12.5 to observe the status of the embryos to determine the pregnancy outcome ( Figure 4 ).

[0043] The results showed that pristane induction caused 37.5% of the embryos in a mouse litter to undergo embryo resorption (no surviving embryos were visible to the naked eye, instead, blood clots absorbed by the mother) or developmental changes (the fetal brain became significantly smaller, indicating abnormal fetal brain development, and the fetal liver turned white, indicating abnormal fetal liver hematopoiesis) ( Figure 4 In the experiment, only 5.4% of embryos in a litter of pristane-induced mice treated with LCA showed embryo resorption or developmental changes ( Figure 4 ). The results showed that LCA treatment could significantly alleviate the embryo resorption or developmental changes induced by pristane.

[0044] On the other hand, in the pristane-induced SLE model, LCA treatment significantly increased the survival rate and body weight of offspring mice ( Figure 6 AD).

[0045] Example 3. In vivo functional experiments demonstrate that LCA can treat and alleviate adverse pregnancy outcomes in spontaneous SLE

[0046] NZW mice are a mouse model of spontaneous SLE and have been reported to have poor pregnancy outcomes.

[0047] For NZW pregnant mice, 10 mg / kg LCA was injected intraperitoneally at E6.5, E9.5, E10.5 and E11.5. For NZW control mice, we did not do any treatment and observed the proportion of embryo absorption in their natural state. The mice were sacrificed and dissected at E12.5, and the embryo status was observed to determine the pregnancy outcome ( Figure 5 ).

[0048] The results showed that 58.3% of the control group mouse embryos had embryo resorption at E12.5, while 28.6% of the LCA-treated group mouse embryos had embryo resorption at E12.5 ( Figure 5 ). The results showed that LCA treatment could significantly improve the survival rate of NZW mouse embryos at E12.5.

[0049] Example 4. Evaluation of the safety and effectiveness of LCA - Effects on offspring survival rate and body weight

[0050] The effect of LCA treatment alone on the health status of offspring mice was evaluated using the method described in Example 1. In the absence of exposure to IFN-I inducers, the survival rate and body weight of offspring mice in the LCA treatment group did not change compared with the solvent control group ( Figure 6 A and B).

[0051] The effect of LCA treatment alone on the health status of offspring mice was evaluated using the method described in Example 2. In the absence of exposure to IFN-I inducers, the survival rate and body weight of offspring mice in the LCA treatment group did not change compared with the solvent control group ( Figure 6 C and D).

[0052] The specific embodiments described above further illustrate the objectives, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above description is only a specific embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. Use of chicoric acid in the preparation of a medicine or kit for preventing, alleviating and / or treating adverse pregnancy outcomes.

2. Use of LCA in the preparation of a drug or kit for preventing, alleviating and / or treating adverse pregnancy outcomes.

3. The method according to claim 1 or 2, wherein the adverse pregnancy outcome is an adverse pregnancy outcome caused by excessive activation of IFN-I, preferably an adverse pregnancy outcome caused by SLE.

4. The method according to claim 3, wherein the IFN-I overactivation is IFN-I overactivation caused by SLE.

5. The method according to any one of claims 1 to 4, wherein the adverse pregnancy outcome is an adverse pregnancy outcome caused by embryo resorption and / or developmental changes.

6. The method according to any one of claims 1 to 5, wherein the adverse pregnancy outcome is selected from the group consisting of spontaneous abortion, intrauterine growth restriction, developmental changes, premature birth, premature rupture of membranes, fetal malformation, neonatal asphyxia, low birth weight and neonatal intensive care unit admission.

7. The use according to any one of claims 1 to 6, characterized in that The medicine or kit further comprises a pharmaceutically acceptable excipient.

8. The use according to any one of claims 1-7, wherein the medicament or kit is administered at E6.5, E9.5, E10.5 and / or E11.5, preferably at E10.5.

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