Use of chlorogenic acid in the preparation of a drug for preventing or treating adverse pregnancy outcomes

Abstract

The present application relates to the technical field of biotechnology, and particularly relates to application of chicoric acid (especially LCA) in preparation of a medicine for preventing, alleviating and / or treating adverse pregnancy outcomes caused by over-activation of IFN-I.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biotechnology, in particular to the application of chicoric acid (especially LCA) in the preparation of a drug for preventing or treating adverse pregnancy outcomes. BACKGROUND

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

[0003] Long-term and excessive activation or induction of IFN-I signal will impair embryonic development, which can be observed in some autoimmune diseases such as systemic lupus erythematosus (SLE) and Aicardi-Goutieres syndrome. Healthy pregnancy requires IFN-I signal, which plays a role in pregnancy recognition and uterine artery remodeling during pregnancy. In contrast, excessive and chronic IFN-I signal will increase the activation of IFN-induced transmembrane proteins (IFITM), thereby inhibiting the formation of placental syncytial trophoblasts (ST), ultimately leading to fetal death.

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

[0005] Cichoric acid (CA) is a natural compound, and its structural formula is: CA has been reported to have antioxidant and anti-inflammatory functions, and has the potential to alleviate the progression of atherosclerosis. L-chicoric acid (LCA) has been reported to be an effective, selective and reversible inhibitor of HIV-1 integrase, which can inhibit the replication of HIV-1 in tissue culture. SUMMARY

[0006] An object of the present application is to provide a safe and effective precision treatment for pregnant patients with excessive activation of IFN-I signal.

[0007] Preferably, the present application provides a safe and effective precision treatment means for SLE pregnancy patients with IFN-I signal over-activation.

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

[0009] Still another object of the present application is to provide that chicoric acid, preferably LCA, can prevent, alleviate and / or treat adverse pregnancy outcomes associated with IFN-I signal over-activation.

[0010] In an embodiment of the present application, LCA is used to verify its therapeutic or alleviating effect on adverse pregnancy outcomes of patients with IFN-I signal over-activation, and therefore those skilled in the art can understand that chicoric acid (CA) can also achieve the same or similar effect.

[0011] In particular, the present application provides the following technical solutions:

[0012] 1. Use of chicoric acid in the preparation of a drug 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 IFN-I over-activation.

[0015] 4. The use according to item 3, wherein the IFN-I over-activation is IFN-I over-activation 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 alteration.

[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 alteration, preterm birth, premature rupture of membranes, fetal malformation, neonatal asphyxia, low birth weight baby and neonatal intensive care unit admission.

[0018] 7. The use according to any one of items 1-6, 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 drug or kit is administered at E6.5, E9.5, E10.5 and / or E11.5, preferably at E10.5.

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

[0021] In the present application, "E+ days" is the number of days of embryonic development after fertilization. Mice are mated, and the next day the tampons are checked to mark the success of mating, and the day of successful mating is E0.5.

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

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

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

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

[0026] Figure 2 A is the actual picture of the pregnancy outcome of LCA treatment of poly(I:C) induction model; B is the statistics of adverse pregnancy outcomes of LCA treatment of poly(I:C) induction model.

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

[0028] Figure 4 A is the actual picture of the pregnancy outcome of LCA treatment of pristane induction model; B is the statistics of adverse pregnancy outcomes of LCA treatment of pristane induction model.

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

[0030] Figure 6 A is the statistics of offspring survival rate of LCA treatment of poly(I:C) induction model, B is the comparison of offspring body weight of each group; C is the statistics of offspring survival rate of LCA treatment of pristane induction model, D is the comparison of offspring body weight of each group. DETAILED DESCRIPTION

[0031] The following examples will facilitate further understanding of the present application by those of ordinary skill in the art, but do not limit the present application in any form.

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

[0033] The mice used in the examples are 8-15 week old C57BL / 6J female and male mice purchased from Shanghai Slac. 8-10 week old C57BL / 6J female mice were mated with C57BL / 6J male mice, and the next day the tampons were checked to use the tampons as the marker of successful mating, and the mating success date was recorded as E0.5. "E+ days" represents the number of days of embryonic development after fertilization. NZW mice are a spontaneous mouse model of systemic lupus erythematosus (SLE) provided by Shanghai Rheumatism Research Institute of Renji Hospital, Shanghai Jiaotong University, which can be constructed by conventional methods or can be commercially available.

[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 experiment demonstrates that LCA can treat and alleviate adverse pregnancy outcomes caused by poly(I:C) induced over-activation of IFN-I

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

[0037] E10.5 and E11.5 mice were injected intraperitoneally 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 state of the embryos to determine the pregnancy outcome Figure 1 ).

[0038] The results show that poly(I:C) induction leads to 44.8% of the embryos in a mouse litter to be resorbed (macroscopically invisible surviving embryos are replaced by blood clots absorbed by the mother) or to develop changes (the fetal brain is significantly smaller, indicating abnormal fetal brain development; the fetal liver is white, indicating abnormal fetal liver hematopoiesis) Figure 2). The results showed that pristane induction caused 37.5% of the embryos in a litter to be resorbed (macroscopically invisible surviving embryos were replaced by blood clots resorbed by the mother) or to have developmental changes (obvious smaller fetal brain suggested abnormal fetal brain development, and white fetal liver suggested abnormal fetal liver hematopoiesis) (Fig. 2A). Figure 2 The results showed that LCA treatment significantly alleviated pristane-induced resorption or developmental changes of embryos.

[0039] On the other hand, in the pristane-induced SLE model, LCA treatment significantly improved the survival rate and body weight of offspring mice (Fig. 2B-D). Figure 6 A-D).

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

[0041] E10.5 mice were injected intraperitoneally with 500 μl of pristane to induce IFN-I signal activation, i.e., to successfully establish a SLE model; 500 μl of normal saline was injected as a control.

[0042] E10.5 mice were injected intraperitoneally with 10 mg / kg of 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 state of the embryos to determine the pregnancy outcome (Fig. 1). Figure 4 ).

[0043] The results showed that pristane induction caused 37.5% of the embryos in a litter to be resorbed (macroscopically invisible surviving embryos were replaced by blood clots resorbed by the mother) or to have developmental changes (obvious smaller fetal brain suggested abnormal fetal brain development, and white fetal liver suggested abnormal fetal liver hematopoiesis) (Fig. 2A). Figure 4 ). The results showed that LCA treatment significantly alleviated pristane-induced resorption or developmental changes of embryos. Figure 4 ).

[0044] On the other hand, in the pristane-induced SLE model, LCA treatment significantly improved the survival rate and body weight of offspring mice (Fig. 2B-D). Figure 6 A-D).

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

[0046] NZW mice are a mouse model of spontaneous SLE, which has 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 the NZW control group of mice, we did not do anything, and observed the proportion of embryo resorption in the natural state. At E12.5, the mice were sacrificed and dissected, and the state of the embryos was observed to determine the pregnancy outcome. Figure 5 ).

[0048] It was found that 58.3% of the embryos of the control group of mice were resorbed at E12.5, and 28.6% of the embryos of the LCA-treated group of mice were resorbed at E12.5 ( Figure 5 ). The results showed that LCA treatment can significantly improve the survival rate of NZW mouse embryos at E12.5.

[0049] Example 4. LCA safety and efficacy evaluation - effect on offspring survival and body weight

[0050] The effect of LCA treatment alone on the health 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-treated group were not changed compared to the solvent control group ( Figure 6 A and B).

[0051] The effect of LCA treatment alone on the health 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-treated group were not changed compared to the solvent control group ( Figure 6 C and D).

[0052] The specific embodiments described above further illustrate the objects, technical solutions and beneficial effects of the present application. It should be understood that the above description is only for specific embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. Use of LCA in the manufacture of a medicament or kit for preventing, alleviating and / or treating an adverse pregnancy outcome, said adverse pregnancy outcome being an adverse pregnancy outcome resulting from IFN-I hyperactivation.

2. Use of LCA in the manufacture of a medicament or kit for preventing, alleviating and / or treating an adverse pregnancy outcome, said adverse pregnancy outcome being an adverse pregnancy outcome resulting from IFN-I hyperactivation.

3. Use of LCA in the manufacture of a medicament or kit for preventing, alleviating and / or treating an adverse pregnancy outcome, said adverse pregnancy outcome being an adverse pregnancy outcome resulting from SLE.

4. Use of LCA in the manufacture of a medicament or kit for preventing, alleviating and / or treating an adverse pregnancy outcome, said adverse pregnancy outcome being an adverse pregnancy outcome resulting from SLE.

5. Use according to claim 1 or 2, wherein said IFN-I hyperactivation is an IFN-I hyperactivation resulting from SLE.

6. Use according to any one of claims 1-4, wherein said adverse pregnancy outcome is an adverse pregnancy outcome resulting from embryo resorption and / or altered development.

7. Use according to any one of claims 1-4, wherein said adverse pregnancy outcome is selected from the group consisting of spontaneous abortion, intrauterine growth restriction, altered development, preterm birth, premature rupture of membranes, fetal malformation, neonatal asphyxia, low birth weight baby and neonatal intensive care unit admission.

8. Use according to any one of claims 1 to 4, characterized in that, said medicament or kit further comprising a pharmaceutically acceptable excipient.

9. Use according to any one of claims 1-4, wherein said medicament or kit is administered at E6.5, E9.5, E10.5 and / or E11.

5.

10. Use according to any one of claims 1-4, wherein said medicament or kit is administered at E10.5.

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