Use of zanamivir in the preparation of a medicament for the treatment or prevention of pre-eclampsia

Abstract

The application provides use of zanamivir in preparation of a medicine for treating or preventing preeclampsia, and a corresponding medicine and treatment method; zanamivir can effectively improve high blood pressure, proteinuria, creatinine, neuraminidase and sialic acid elevation symptoms of preeclampsia, increase NO level, and improve pregnancy outcome. Zanamivir and hydroxyprogesterone caproate in combination show better effects.

Description

TECHNICAL FIELD

[0001] The present application provides the use of zanamivir in the manufacture of a medicament for the treatment or prevention of pre-eclampsia, as well as the corresponding medicaments and methods of treatment. BACKGROUND

[0002] Pre-eclampsia, also known as eclampsia or toxemia of pregnancy, is a clinical syndrome that occurs after 20 weeks of gestation and is characterized by hypertension, proteinuria and oedema, and associated symptoms such as headache, visual disturbances, nausea, vomiting, abdominal discomfort and others. Pre-eclampsia not only affects the development of the foetus, but also often endangers the life of the pregnant woman. The pathophysiology of the disease is generally divided into two stages: 1) abnormal placental development, particularly vasodilatory dysfunction of the uterine spiral arterioles, and the impact on maternal blood flow to the foetus. Reduced placental perfusion activates the placenta to release associated cytokines and causes systemic haemodynamic changes. 2) The pregnant woman develops clinical symptoms, mainly circulatory disorders caused by maternal vascular endothelial cell dysfunction, including changes in vascular reactivity, activation of the coagulation cascade and disruption of vascular integrity. Pre-eclampsia affects most organs of the pregnant woman, but mainly the kidneys, liver and brain.

[0003] Although there are a large number of preclinical and clinical studies on preeclampsia, its pathogenesis is currently known to be complex but not clear, and the corresponding treatment means is also quite scarce, generally only for symptomatic antihypertensive, diuretic, sedative treatment, etc., severe eclampsia can use magnesium sulfate to relieve spasm symptoms, and currently the most effective clinical treatment measure is to terminate pregnancy. The etiology of preeclampsia may be that on the basis of multiple genetic factors, susceptible factors lead to abnormal cellular immunity, placental and trophoblast ischemia and hypoxia, abnormal oxidative stress response, causing secondary systemic vascular endothelial cell damage and leading to the occurrence of the disease. In view of this, the scientific community has proposed a series of mechanism theories including placental or trophoblast ischemia theory, oxidative stress theory, immune regulation abnormality theory, genetic theory and vascular endothelial damage theory to explain the pathogenesis of preeclampsia. Placental spiral arteriole remodeling disorders and placental shallow implantation, trophoblast ischemia and hypoxia and vascular endothelial cell damage are generally considered to be the pathophysiological basis of preeclampsia. A large number of studies have shown that the release of placental and vascular-derived circulating factors due to ischemia and hypoxia, including vascular endothelial growth factor (VEGF), soluble vascular endothelial growth factor-1 (sFlt-1), anti-angiotensin II receptor 1 autoantibodies and tumor necrosis factor-α (TNF-α), interleukin 1 (IL-1) and IL-6, pentraxin 3 (PTX3) and the like can cause maternal vascular endothelial dysfunction through different pathways. In addition, the proportion of vasodilator and vasoconstrictor in the blood of patients with preeclampsia is imbalanced, and the vascular endothelium is damaged, which increases the sensitivity of blood vessels to vasoconstrictors and decreases the sensitivity to vasodilators, thereby causing systemic arteriolar spasm. Patients with pre-existing hypertension, diabetes, obesity and hyperlipidemia may have oxidative stress and vascular endothelial dysfunction before pregnancy, and are more likely to develop preeclampsia after pregnancy.

[0004] Vascular endothelial cells are directly exposed to the shear stress and are the responders of the shear stress. Endothelial cells convert the extracellular mechanical signals into intracellular signals, and then trigger downstream biochemical reactions. The glycocalyx is a layer of brush-like polysaccharide-protein complex structure on the apical membrane of endothelial cells, which is located between the vessel wall and blood, is a dynamic natural barrier on the surface of endothelial cells, and mediates the force transmission function of endothelial cells. The vascular endothelial glycocalyx is a highly negatively charged surface layer of endothelial cells composed of oligosaccharide chain glycoproteins containing sialic acid residues at the end and proteoglycans containing glycosaminoglycan side chains. In addition to being a selective permeability barrier of the vascular wall, regulating the rheological properties of the microcirculation, and participating in vascular protection and signal transmission, a large amount of evidence shows that the endothelial glycocalyx is also involved in the sensing and transmission of the flow shear stress. Manolis et al. treated bovine thoracic aortic endothelial cells by simultaneously selectively degrading sialic acid, heparin sulfate, chondroitin sulfate and hyaluronic acid in the glycocalyx by using selective enzyme degradation method, and found that the synthesis of nitric oxide (NO) induced by flow shear stress was blocked, indicating that the intact glycocalyx plays an important role in the synthesis of NO induced by shear stress. Studies have also shown that when the glycocalyx is intact, the flow shear stress can be transmitted to the actin cytoskeleton through the core protein, or directly transmitted to the cell membrane through the core protein, thereby mediating the downstream signal transduction of the cell, such as the generation of NO and the reorganization of the cytoskeleton. When the glycocalyx is damaged, the normal function of vascular endothelial cells is affected. A recent clinical study found that more glycocalyx degradation can be detected in patients with early-onset preeclampsia, and is accompanied by an increase in the levels of heparin sulfate proteoglycans and hyaluronic acid in the blood, thereby indicating that the degradation of vascular endothelial glycocalyx is closely related to the onset of early-onset preeclampsia.

[0005] The degradation of the vascular endothelial glycocalyx is usually mediated by a variety of specific enzymes. Neuraminidase, also known as sialidase, can act on oligosaccharide chain glycoproteins containing sialic acid residues at the end of the glycocalyx, hydrolyze the ketoside bond to release sialic acid, and ultimately cause glycocalyx degradation and corresponding impairment of vascular endothelial function. Some documents have shown that the level of neuraminidase can be detected in the blood of patients with some cardiovascular system-related diseases such as type II diabetes, and inhibiting neuraminidase can improve the function of vascular endothelial cells and the whole cardiovascular system by inhibiting glycocalyx degradation. In view of the fact that there is no effective drug for preventing and treating preeclampsia in the clinic at present, there is a great need for drugs for preventing and treating the disease in the art, and drugs for inhibiting neuraminidase provide a new idea and means for the development of drugs for preventing and treating preeclampsia. SUMMARY

[0006] In one aspect, the present application provides the use of zanamivir in the preparation of a medicament for treating or preventing preeclampsia.

[0007] The zanamivir includes the compound zanamivir and its medically or pharmaceutically acceptable derivatives or salts.

[0008] Further, the pharmaceutical dosage form is an injection, oral preparation or external preparation; preferably an injection, oral preparation; particularly preferably an injection.

[0009] Further, the injection includes an injection solution, a powder injection; the oral preparation includes a tablet, a capsule, an oral solution, a throat or nasal spray, and the external preparation includes an ointment, a spray, a patch.

[0010] Further, the treatment or prevention of preeclampsia is to improve headache, blurred vision, malaise, vomiting, abdominal discomfort, hypertension, proteinuria, edema, liver and kidney function, thrombocytopenia and / or coagulation system abnormalities; creatinine, neuraminidase and sialic acid elevation, and blood NO level reduction; and to improve pregnancy outcome.

[0011] In another aspect, the present application provides a method for treating or preventing preeclampsia, comprising administering zanamivir to a patient at risk of or suffering from preeclampsia.

[0012] Further, the pharmaceutical dosage form is an injection, oral preparation or external preparation; preferably an injection, oral preparation; particularly preferably an injection.

[0013] The injection includes an injection solution, a powder injection; the oral preparation includes a tablet, a capsule, an oral solution, a throat or nasal spray, and the external preparation includes an ointment, a spray, a patch.

[0014] Further, the treatment or prevention of preeclampsia is to improve headache, blurred vision, malaise, vomiting, abdominal discomfort, hypertension, proteinuria, edema, liver and kidney function, thrombocytopenia and / or coagulation system abnormalities; creatinine, neuraminidase and sialic acid elevation, and blood NO level reduction; and to improve pregnancy outcome.

[0015] Further, the administration method of zanamivir can be selected from intramuscular injection, intravenous injection, intravenous infusion, preferably intramuscular injection; 1-7 times per week, preferably 1-4 times; more preferably 1-2 times; the duration of administration is 1-30 days, preferably 1-7 days, more preferably 1-7 days.

[0016] In another aspect, the present application provides a drug for treating or preventing preeclampsia.

[0017] Further, the pharmaceutical dosage form is an injection, oral preparation or external preparation; preferably an injection, oral preparation; particularly preferably an injection; further preferably an intramuscular injection.

[0018] Further, the medicine comprises instructions, wherein the instructions indicate that the method of administration of zanamivir is 1-7 times per week, preferably 1-4 times per week, more preferably 1-2 times per week; and the duration of administration is 1-30 days, preferably 1-7 days, more preferably 1-7 days.

[0019] Further, the treatment or prevention of preeclampsia is to improve headache, blurred vision, nausea, vomiting, abdominal discomfort, hypertension, proteinuria, edema, liver and kidney function, thrombocytopenia and / or coagulation system abnormalities; creatinine, neuraminidase and sialic acid increase, blood NO level decrease; and improve pregnancy outcome.

[0020] Further, the method or medicine of the present application has a dosage of 20-40 mg / day, 90 mg / kg or 120 mg / kg, preferably 20 mg / day, when administered.

[0021] Further, the medicine further comprises a pharmaceutically acceptable excipient, which includes but is not limited to solvents, cosolvents, stabilizers, dispersants, viscosity regulators, antioxidants, sweeteners, binders, gas generating agents.

[0022] Further, zanamivir is the only effective component in the method or medicine; or the method further comprises administration of other effective components or the medicine further comprises other effective components; the other effective components include but are not limited to antihypertensive drugs, protein supplements, anti-dizziness drugs, cardiovascular disease drugs.

[0023] In another aspect, the present application provides a neuraminidase inhibitor for use in the preparation of a medicine for treating or preventing preeclampsia.

[0024] The neuraminidase inhibitor includes a compound neuraminidase inhibitor and its medically or pharmaceutically acceptable derivative or salt.

[0025] Further, the medicine dosage form is an injection, an oral preparation or a topical preparation; preferably an injection, an oral preparation; particularly preferably an injection.

[0026] Further, the injection includes an injection solution, a powder injection; the oral preparation includes a tablet, a capsule, an oral solution, a throat or nasal spray, and the topical preparation includes an ointment, a spray, a patch.

[0027] Further, the treatment or prevention of preeclampsia is to improve headache, blurred vision, nausea, vomiting, abdominal discomfort, hypertension, proteinuria, edema, liver and kidney function, thrombocytopenia and / or coagulation system abnormalities; creatinine, neuraminidase and sialic acid increase, blood NO level decrease; and improve pregnancy outcome.

[0028] In another aspect, the present application provides a method for treating or preventing preeclampsia, comprising administering a neuraminidase inhibitor to a patient at risk of or suffering from preeclampsia.

[0029] Further, the neuraminidase inhibitor is in the form of an injection, an oral preparation or a topical preparation; preferably an injection, an oral preparation; particularly preferably an injection.

[0030] The injection includes an injection solution, a powder injection; the oral preparation includes a tablet, a capsule, an oral solution, a throat or nasal spray; the topical preparation includes an ointment, a spray, a patch.

[0031] Further, the treatment or prevention of preeclampsia is to improve headache, blurred vision, malaise, vomiting, abdominal discomfort, hypertension, proteinuria, edema, liver and kidney function, thrombocytopenia and / or coagulation system abnormalities; creatinine, neuraminidase and sialic acid elevation, blood NO level reduction; and improve pregnancy outcome.

[0032] Further, the method of administration of the neuraminidase inhibitor can be selected from intramuscular injection, intravenous injection, intravenous infusion, preferably intramuscular injection; 1-7 times per week, preferably 1-4 times; more preferably 1-2 times; the duration of administration is 1-30 days, preferably 1-7 days, more preferably 1-7 days.

[0033] In another aspect, the present application provides a drug for treating or preventing preeclampsia.

[0034] Further, the drug is in the form of an injection, an oral preparation or a topical preparation; preferably an injection, an oral preparation; particularly preferably an injection; further preferably an intramuscular injection.

[0035] Further, the drug comprises instructions, which record the method of administration of the neuraminidase inhibitor as 1-7 times per week, preferably 1-4 times; more preferably 1-2 times; the duration of administration is 1-30 days, preferably 1-7 days, more preferably 1-7 days.

[0036] Further, the treatment or prevention of preeclampsia is to improve headache, blurred vision, malaise, vomiting, abdominal discomfort, hypertension, proteinuria, edema, liver and kidney function, thrombocytopenia and / or coagulation system abnormalities; creatinine, neuraminidase and sialic acid elevation, blood NO level reduction; and improve pregnancy outcome.

[0037] Further, the method or drug of the present application has a dosage of 20-40 mg / day, 90 mg / kg or 120 mg / kg, preferably 20 mg / day, when administered.

[0038] Further, the medicine also comprises pharmaceutically acceptable adjuvants, which include but are not limited to solvents, co-solvents, stabilizers, dispersants, viscosity regulators, antioxidants, sweeteners, binders, gas generating agents.

[0039] Further, the neuraminidase inhibitor is the only effective component in the method or medicine; or in the method, other effective components are also administered or in the medicine, other effective components are also contained; the other effective components include but are not limited to hypotensive drugs, protein supplements, anti-dizziness drugs, cardiovascular disease drugs.

[0040] The neuraminidase inhibitor in the present application is not limited to the chemical structure of CAS No. 139110-90-8 (i.e., 5-acetylamino-4-[(aminoiminomethyl)-amino]-2,6-hydro-3,4,5-trideoxy-D-glycero-D-galacto-2-enonic acid, or 4-guanidino-2-deoxy-2,3-didehydro-N-acetylneuraminic acid), but also includes its medically or pharmaceutically acceptable derivatives or salts. The neuraminidase inhibitor in the present application refers to a compound or composition that inhibits neuraminidase, including various neuraminidase inhibitors known and researched in the art, including but not limited to zanamivir.

[0041] The hydroxyprogesterone caproate in the present application is also known as 17-alpha hydroxyprogesterone caproate, 17-HPC, 17-hydroxyprogesterone caproate, 17-hydroxyprogesterone caproate, CAS No. 630-56-8.

[0042] The preeclampsia in the present application refers to a clinical syndrome during pregnancy, which is mainly characterized by high blood pressure, proteinuria and edema, accompanied by headache, blurred vision, nausea, vomiting, abdominal discomfort, etc. The main symptoms also include elevated creatinine, neuraminidase and sialic acid, and reduced blood NO levels, and the specific symptoms include but are not limited to the above.

[0043] The zanamivir in the present application can be administered as the only effective component for treating preeclampsia or prepared into a corresponding medicine, or can be administered in combination with known or researched drugs for treating preeclampsia or symptoms associated with preeclampsia, or prepared into a drug combination or a pharmaceutical composition. These drugs include but are not limited to hypotensive drugs, protein supplements, anti-dizziness drugs, cardiovascular disease drugs, etc.

[0044] The zanamivir in the present application can be administered by intramuscular injection, intravenous injection, intravenous infusion, etc., and is preferably administered by intramuscular injection.

[0045] Depending on the dosage form and the condition, the zanamivir in the present application can be administered at a frequency of 1-7 times per week, preferably 1-4 times, and more preferably 1-2 times; and the duration of administration can be 1-30 days, preferably 1-14 days, and more preferably 1-7 days.

[0046] The amount of zanamivir to be administered in the present application is not limited to the range described in the specification and claims, and the clinician can adjust the amount according to the patient's condition and the drug dosage form and the like.

[0047] The dosage form of zanamivir of the present application includes, but is not limited to, injections such as injection solutions, powder injections and the like, oral preparations such as tablets, capsules, oral solutions and the like, throat or nasal sprays, external preparations such as ointments, sprays, patches and the like; injections, oral preparations are preferred; injections are particularly preferred.

[0048] The pregnancy outcomes described in the present application include, but are not limited to, miscarriage and fetal / infant health problems related to pre-eclampsia or blood pressure.

[0049] The person skilled in the art can select suitable excipients for the drugs involved in the present application according to general knowledge in the pharmaceutical field, and the available excipient categories include, but are not limited to, solvents, cosolvents, stabilizers, dispersants, viscosity modifiers, antioxidants, sweeteners, binders, gas generating agents and the like.

[0050] In the present application, the amount of neuraminidase inhibitor such as zanamivir to be administered can be 90 mg / kg-120 mg / kg; preferably 120 mg / kg.

[0051] The experimental results show that zanamivir can improve the symptoms of hypertension and proteinuria in pre-eclampsia and reduce the creatinine level in urine; zanamivir can reduce the levels of neuraminidase and sialic acid in blood and increase the level of NO in blood. Zanamivir combined with hydroxyprogesterone caproate shows better preventive and therapeutic effects on pre-eclampsia. It provides a brand-new idea and means for the development of drugs for preventing and treating pre-eclampsia. BRIEF DESCRIPTION OF DRAWINGS

[0052] Figure 1 Figure 4 is a graph showing the mean arterial pressure of pregnant rats in each group on the 12th day of pregnancy; it shows that zanamivir (120 mg / kg) can significantly reduce the increase in mean arterial pressure of pregnant rats caused by L-NAME (data are mean ± standard error, N = 9-22, * p < 0.05);

[0053] Figure 2 Figure 5 is a graph showing the diastolic pressure of pregnant rats in each group on the 12th day of pregnancy; it shows that zanamivir (120 mg / kg) can significantly reduce the increase in diastolic pressure of pregnant rats caused by L-NAME (data are mean ± standard error, N = 9-22, * p < 0.05);

[0054] Figure 3Figure 3 is a graph showing systolic blood pressure in pregnant rats on day 12 of pregnancy for each group; it shows that zanamivir (120 mg / kg) can significantly reduce the increase in systolic blood pressure in pregnant rats induced by L-NAME (data are mean ± SEM, N = 9-22, **p<0.01);

[0055] Figure 4 Figure 4 is a graph showing 24-hour urine protein in pregnant rats on days 17-18 of pregnancy for each group; it shows that zanamivir can significantly reduce the increase in 24-hour urine protein in pregnant rats induced by L-NAME (data are mean ± SEM, N = 5-22, **p<0.01, ***p<0.001);

[0056] Figure 5 Figure 5 is a graph showing the number of pups born by pregnant rats for each group; zanamivir can significantly improve the decrease in the number of pups born by pregnant rats induced by L-NAME (data are mean ± SEM, N = 9-16, *p<0.05). DETAILED DESCRIPTION

[0057] Example 1: Establishment of an animal model of preeclampsia

[0058] Pregnant CD-1 mice (age 8-10 weeks) were randomly divided into a control group and a preeclampsia model group. From the 7th to 16th day of pregnancy, the model group of mice was induced to develop preeclampsia by subcutaneous injection of 50 mg / kg of N-nitro-L-arginine methyl ester (L-NAME) every day. The control group was subcutaneously injected with the same volume of solvent.

[0059] Example 2: Experimental grouping

[0060]

[0061]

[0062] Example 3: Evaluation of therapeutic effect

[0063] The L-NAME-induced preeclampsia animal model, according to the literature reports, will develop hypertension symptoms two days after the first administration of L-NAME and will last at least five days after the first administration of L-NAME. The appearance of proteinuria symptoms is relatively late. Therefore, in this experiment, we focused on monitoring the blood pressure changes of pregnant rats on the 12th day of pregnancy (i.e., five days after the first administration of L-NAME) and collecting 24-hour urine on the 17th-18th day of pregnancy to determine the total protein level. At the same time, we also recorded the number of pups produced by pregnant rats in each group.

[0064] Example 4: Experimental results

[0065] The results of blood pressure testing in pregnant rats on the 12th day of pregnancy showed that zanamivir (120 mg / kg) can significantly reduce the increase in systolic blood pressure in pregnant rats induced by L-NAME (data are mean ± SEM, N = 9-22, **p<0.01); Figures 1-3), and the mean arterial pressure, systolic pressure and diastolic pressure of the pregnant model group were significantly higher than that of the pregnant control group, indicating that this animal model successfully showed the hypertension symptoms of preeclampsia. Among them, 90mg / kg zanamivir had no significant change on blood pressure. However, 120mg / kg zanamivir administration could significantly reduce the increase of mean arterial pressure, systolic pressure and diastolic pressure of pregnant mice caused by L-NAME, and all had statistically significant differences.

[0066] The total protein in 24-hour urine of the 17-18 day pregnant mice was determined. Figure 4 ), and the 24-hour urine protein of the pregnant model group was significantly higher than that of the pregnant control group, indicating that this animal model successfully showed the proteinuria symptoms of preeclampsia. Zanamivir administration at 90mg / kg and 120mg / kg could significantly reduce the increase of urine protein of pregnant mice caused by L-NAME, and all had statistically significant differences.

[0067] In this experiment, we also recorded the number of pups produced by each group of pregnant mice. As shown in Figure 5 , compared with the pregnant control group, the number of pups produced by the pregnant model group was significantly reduced. Zanamivir administration at 90mg / kg and 120mg / kg could significantly improve the reduction of the number of pups produced by pregnant mice caused by L-NAME, and the number of pups in the 120mg / kg administration group returned to the level comparable to the pregnant control group, and had statistically significant differences.

[0068] In summary, zanamivir can significantly improve the hypertension and proteinuria symptoms of L-NAME-induced preeclampsia in pregnant mice, and increase the number of pups produced by pregnant mice. The results suggest that zanamivir has a preventive and therapeutic effect on preeclampsia, can improve the maternal hypertension and proteinuria symptoms, and also has the potential to protect fetal development.

Claims

1. Use of zanamivir in the preparation of a medicament for the treatment or prevention of pre-eclampsia.

2. Use according to claim 1, wherein the pharmaceutical dosage form is an injection, oral preparation.

3. Use according to claim 2, wherein the injection is an injection solution or powder injection; the oral preparation is a tablet, capsule or oral solution.

4. Use according to any one of claims 1 to 3, wherein the treatment or prevention of pre-eclampsia is to improve hypertension and / or proteinuria and to improve pregnancy outcome.

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