Application of L-malic acid in preparation of medicine for treating intrahepatic cholestasis in gestation period

Abstract

The invention discloses application of L-malic acid in preparation of a medicine for treating intrahepatic cholestasis in a gestation period, and belongs to the field of medicine. It is found that L-malic acid can improve intrahepatic cholestasis in the gestation period, relieve liver injury and reduce serum total bile acid, and then the gestation outcome is improved. A new treatment means is provided for preparing the medicine for treating the intrahepatic cholestasis in the gestation period, the L-malic acid is applied to the development process of the medicine related to the intrahepatic cholestasis in the gestation period, and a new means and approach are provided for treating the intrahepatic cholestasis in the gestation period.

Description

Technical Field

[0001] The present invention relates to the application of L-malic acid in the preparation of a drug for treating intrahepatic cholestasis of pregnancy, and belongs to the field of medicine. Background Art

[0002] Intrahepatic cholestasis of pregnancy (ICP) is a liver disease that occurs only during pregnancy, characterized by symptoms such as pruritus, jaundice, and elevated bile acids, and has a high recurrence rate in subsequent pregnancies. ICP increases the risk of maternal hepatobiliary diseases and can lead to adverse perinatal outcomes, including fetal ischemia and hypoxia, spontaneous preterm birth, neonatal respiratory distress, and stillbirth.

[0003] Ursodeoxycholic acid (UDCA) is a natural hydrophilic bile acid, accounting for only 3% - 5% of human physiological bile acids. It was originally used to treat cholesterol gallstones, cholestatic liver diseases, and other hepatobiliary diseases. International and Chinese Medical Association ICP-related guidelines both recommend ursodeoxycholic acid (UDCA) as the first-choice drug for treating ICP. However, its therapeutic effect is not good [Lancet. 2019 Sep 7; 394(10201): 849 - 860].

[0004] The glutathione precursor S-adenosyl-methionine (SAMe) is also a drug widely used in the clinical treatment of ICP. SAMe affects the composition and fluidity of the hepatocyte plasma membrane, enhancing the methylation of hormone metabolites and bile excretion. The 2015 edition of the Chinese Medical Association guidelines recommends SAMe as a second-line drug or combination therapy drug for ICP, with a dose of 1 g / d by intravenous drip for a course of 12 - 14 days; 500 mg orally, twice a day. Similar to UDCA, there is currently no evidence that SAMe has a definite therapeutic effect on treating ICP and can improve pregnancy outcomes [Cochrane database of systematic reviews 2020 Jul 27; 7(7)].

[0005] Therefore, it is urgent to search for and screen effective drugs for treating ICP. Summary of the Invention

[0006] To solve the above problems, the present invention discovers that the use of L-malic acid (MA) can relieve ICP liver injury and improve pregnancy outcomes.

[0007] MA is an intermediate of the tricarboxylic acid cycle, and its dietary sources are immature fruits, including immature hawthorns, apples, and grapes. Since the content of MA in fruits is low, MA is mainly produced by enzymatic synthesis and microbial fermentation. Currently, there is no research report on the role of MA in ICP.

[0008] The first object of the present invention is to provide the use of MA or a composition containing MA in the preparation of a drug for treating ICP.

[0009] In one embodiment, the symptoms of intrahepatic cholestasis of pregnancy are manifested as itching and an increase in the total bile acid level.

[0010] In one embodiment, the drug is any pharmaceutically acceptable dosage form.

[0011] In one embodiment, the dosage forms include: tablets, inhalation preparations, oral liquids, injections, and topical preparations.

[0012] In one embodiment, the composition further contains excipients.

[0013] In one embodiment, the excipients are pharmaceutically acceptable excipients; optionally, the excipients include one or more of probiotics, lactose, maltose, microcrystalline cellulose, carboxymethyl cellulose, hydroxypropyl methylcellulose, starch, sucrose, glucose, aspartame, water, glycerol, whey protein powder, and chitosan oligosaccharide.

[0014] The second object of the present invention is to provide a pharmaceutical composition for treating intrahepatic cholestasis of pregnancy, and the pharmaceutical composition contains MA.

[0015] In one embodiment, the pharmaceutical composition further includes one or more of S-adenosylmethionine, dexamethasone, Yinchenhao Decoction, Transmetil, Ursofalk, ademetionine 1,4-butanedisulfonate, Yinzhihuang Oral Liquid, and probiotics.

[0016] In one embodiment, the probiotics include one or more of Bifidobacterium, Lactobacillus, and Bacillus subtilis.

[0017] The third object of the present invention is to provide the use of MA or a composition containing MA in the preparation of a drug for improving the placental function of intrahepatic cholestasis of pregnancy and promoting vascular development.

[0018] Advantages of the present invention

[0019] To solve the above problems, the present invention discovers that the use of MA can improve ICP and relieve liver injury, and on this basis, improve pregnancy outcomes. The present invention provides a new treatment method for preparing drugs for treating ICP, applying MA to the drug development process related to ICP to better treat ICP. The present invention provides a brand-new idea for the current drugs for treating ICP, broadens the selection field of drugs for treating ICP, and has important application and popularization significance.

[0020] Specifically:

[0021] (1) MA can significantly inhibit the abnormal increase of TBA in ICP mice;

[0022] (2) MA can significantly improve liver injury in ICP mice and inhibit the abnormal increase of ALT and AST in serum;

[0023] (3) MA can significantly improve the placental function of mice, increase the placental weight, and reduce the placental / fetal weight ratio. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a schematic diagram of MA reducing the content of TBA in mice, with TBA in serum, liver, and placental tissues as indicators.

[0025] Figure 2 It is a schematic diagram of MA improving the liver function of mice, with liver ALT, AST, and H&E staining as indicators.

[0026] Figure 3 It is a schematic diagram of MA improving the placental function of mice, with placental weight, fetal weight, and placental / fetal weight ratio as indicators.

[0027] Figure 4 It is a schematic diagram of MA promoting the placental vascular development of ICP mice, with the blood sinus area in H&E staining as an indicator. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0028] The following will clearly and completely describe the concept and technical effects of the present invention in combination with the embodiments to fully understand the purpose, features, and effects of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative efforts shall fall within the protection scope of the present invention.

[0029] Raw materials:

[0030] L-Malic acid is purchased from: Wuxi Fanbo Biotechnology Co., Ltd.;

[0031] C57BL / 6 mice are all from the Experimental Animal Center of Jiangnan University.

[0032] Detection method:

[0033] 1. Biochemical index detection

[0034] Use an alanine aminotransferase detection kit (Nanjing Jiancheng Company) to detect the content of ALT in liver tissue;

[0035] Use an aspartate aminotransferase detection kit (Nanjing Jiancheng Company) to detect the content of AST in liver tissue;

[0036] Use a total bile acid detection kit (Nanjing Jiancheng Company) to detect the content of TBA in serum, placenta and liver tissue.

[0037] 2. Histopathological injury detection

[0038] First, soak the mouse placenta and liver tissues in 4% paraformaldehyde for fixation, dehydrate them with gradient alcohol with concentrations from 70% to 100%, clear the tissues with 100% xylene after dehydration, then immerse them in paraffin in a wax bath, embed the tissues, and finally make paraffin sections. After staining with an H&E staining kit (Nanjing Jiancheng Company), observe and take pictures under a microscope.

[0039] Example 1: Application of L - malic acid in the treatment of ICP

[0040] Detect the therapeutic effect of L - malic acid on ICP through animal experiments. The specific method is as follows:

[0041] 1. Animal experiment

[0042] Use 6 - 8 - week - old C57BL / 6 mice. Cage adult mice in cages according to a male - female ratio of 2:1. Check the vaginal plugs of female mice the next morning as the standard for successful pregnancy.

[0043] Starting from gestational day 12.5 (GD12.5), female mice are mainly divided into three groups according to their diet for 7 days, namely:

[0044] Normal: Standard maintenance or breeding diet (CRM);

[0045] ICP: CRM + 0.5% cholic acid (CA, add 0.5% cholic acid to normal food) to induce ICP;

[0046] MA + ICP: CRM + 0.5% CA + 2% MA (add 2% MA and 0.5% cholic acid to normal food).

[0047] After 7 days, sacrifice the mice, and collect mouse serum, liver tissue, placenta tissue and fetal mice for biochemical index detection.

[0048] 2. Detection of Therapeutic Effects

[0049] (1) TBA Content

[0050] The TBA content in the serum, liver tissue, and placental tissue of mice was detected, and the results are as Figure 1 shown in Table 1

[0051] The results showed that compared with the normal group, the serum TBA in the model group (ICP) increased by 146%, the TBA in the liver tissue increased by 57%, and the TBA in the placental tissue increased by 68%; while in the treatment group administered with L - malic acid (MA + ICP), compared with the model group, the serum TBA in mice decreased by 46%, the TBA in the liver tissue decreased by 33%, and the TBA in the placental tissue decreased by 41%. That is, L - malic acid has the effect of inhibiting the abnormal increase of TBA in ICP.

[0052] Table 1 TBA Content

[0053]

[0054] (2) Liver Function

[0055] The ALT / AST index is an important biomarker for predicting the liver function of ICP. The ALT and AST values in the liver tissue were detected, and the results are as Figure 2 shown in Table 2

[0056] The results showed that in the liver tissue of mice in the model group (ICP), ALT increased by 43% and AST increased by 48%. The liver H&E staining showed uneven arrangement of hepatic cords, nuclear pyknosis, and vacuolization of cells; while in the treatment group administered with L - malic acid (MA + ICP), compared with the model group, ALT in the liver tissue of mice decreased by 29%, AST decreased by 37%, and the liver H&E staining showed uniform arrangement of hepatic cords, reduced cell vacuolization and nuclear pyknosis. That is, L - malic acid has the effect of improving liver damage in ICP.

[0057] Table 2 Liver Function

[0058]

[0059] (3) Placental Function

[0060] The weight of the mouse placenta, the weight of the fetal mouse, and the ratio of the weight of the placenta to the fetal mouse were detected, and the results are as Figure 3 shown in Table 3

[0061] The results showed that in the model group (ICP), the placental weight of mice decreased by 31%, the fetal weight decreased by 58%, and the placental-fetal weight ratio increased by 73%; while in the treatment group administered with L-malic acid (MA+ICP), compared with the model group, the placental weight increased by 27%, the fetal weight increased by 95%, and the placental-fetal weight ratio decreased by 38%. That is, L-malic acid has the effect of improving placental function in ICP.

[0062] Table 3 Placental function

[0063]

[0064] (4) Placental vascular development

[0065] The blood sinus area in the placenta of mice was detected, and the results are as Figure 4 shown in Table 4.

[0066] The results showed that in the model group (ICP), the blood sinus area of mice decreased by 67%; while in the treatment group administered with L-malic acid (MA+ICP), compared with the model group, the blood sinus area increased by 66%. That is, L-malic acid has the effect of improving placental blood vessels in ICP.

[0067] Table 4 Blood sinus area

[0068]

[0069] Although the present invention has been disclosed above with preferred embodiments, it is not intended to limit the present invention. Anyone familiar with this technology can make various modifications and decorations without departing from the spirit and scope of the present invention. Therefore, the protection scope of the present invention should be defined by the claims.

Claims

1. Use of L-malic acid or a composition containing L-malic acid in the preparation of a medicament for treating intrahepatic cholestasis of pregnancy.

2. The use according to claim 1, characterized in that: Symptoms of intrahepatic cholestasis of pregnancy include pruritus and elevated total bile acid levels.

3. The use according to claim 1, characterized in that: The drug is in any pharmaceutically acceptable dosage form.

4. The use according to claim 3, characterized in that: The dosage forms include: tablets, inhalation preparations, oral liquids, injections, and external preparations.

5. The use according to claim 1, characterized in that: The composition further contains auxiliary materials.

6. The use according to claim 5, characterized in that: The excipients are pharmaceutically acceptable excipients; optionally, the excipients include: one or more of probiotics, lactose, maltose, microcrystalline cellulose, carboxymethyl cellulose, hydroxypropyl methyl cellulose, starch, sucrose, glucose, aspartame, water, glycerol, whey protein powder, and chitosan oligosaccharide.

7. A pharmaceutical composition for treating intrahepatic cholestasis of pregnancy, characterized in that: The pharmaceutical composition contains L-malic acid.

8. The pharmaceutical composition according to claim 7, characterized in that The pharmaceutical composition further comprises: one or more of S-adenosylmethionine, dexamethasone, Yinchenhao decoction, Sumetai, Yousifu, adenosylmethionine butadiene sulfonate, Yinzhihuang oral liquid, and probiotics.

9. The pharmaceutical composition according to claim 8, characterized in that Probiotics include: One or more of Bifidobacterium, Lactobacillus and Bacillus subtilis.

10. Use of L-malic acid or a composition containing L-malic acid in the preparation of a drug for improving placental function in intrahepatic cholestasis of pregnancy and promoting vascular development.

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