NEW SYNTHESIS OF AKETAMINOPHENE COMPOUND WITHOUT SIDE EFFECTS ON THE LIVER

Abstract

A novel compound complex that has no side effects on a liver and is used to alleviate the toxicity of an acetaminophen (APAP) drug to the liver. The composition of the compound consists of (a) acetaminophen in a pharmaceutically effective amount and (b) a commonly-used safe and pharmaceutically acceptable excipient that can be combined with one or more of two drugs to reduce the toxicity of a drug metabolized by the hepatic enzyme CYP2E1 in the liver.The compound is selected from the following group: Tween 20, microcrystalline cellulose, dicalcium phosphate, polyoxyethylene 23 lauryl ether, saccharin, mannitol, polyoxyethylene alkyl ether, sucralose, pyrrolidone, sodium starch glycolate, S100 acrylic resin, sodium carboxymethylcellulose, polyoxyethylene polyoxyethylene, menthol, low-substituted propylcellulose hydrocarbon, pregelatinized starch, Dextrates NF hydrate, citric acid, polyoxyethylene castor oil, colloidal silicon, aliphatic polyethylene glycol monostearate ester, sorbic acid, lemon oil, hydroxypropylcellulose, sorbitol, acesulfame potassium, hypromellose phthalate, lactose monohydrate, maltodextrin, Brij 58, Brij 76, Tween 80, Tween 40, PEG 400, Peg 4000, PEG 2000, and the like, so as to reduce the side effects caused by acetaminophen on the liver.

Description

[0001] New compound combination of acetaminophen without hepatic side effects Technical field

[0002] The present invention relates to a new compound combination of paracetamol (Acetaminophen, APAP) without hepatic effects, in particular to a combination of paraacetaminophen and its ability to inhibit cytochrome P450 2E1 (CYP2E1) enzyme activity. A new compound of acetaminophen, which is a safe and pharmaceutically acceptable common excipient or any combination thereof, to reduce side effects such as liver toxicity caused by acetaminophen. Background technique

[0003] Acetaminophen, also known as paracetamol or N-acetyl-para-aminophenol, APAP for short, is the most commonly used antipyretic analgesic on the market, and many people often use APAP every year Cases of inappropriate poisoning or suicide have occurred. Liver damage caused by APAP is the most important factor causing severe illness and death. Many clinical experiences have proved that the hepatotoxicity of APAP can be prevented. Early diagnosis and real-time administration of the antidote N-acetylcysteine ​​(NAC for short) can effectively prevent the occurrence of hepatotoxicity.

[0004] Early detection of acetaminophen overdose is warranted because the best prognosis is achieved when the antidote is administered within 8 hours of intoxication. The early signs of drug poisoning include physical discomfort, nausea and vomiting, but in some patients, the blood concentration of acetaminophen has reached the toxic level and the liver function is also obviously abnormal, and there are still no symptoms in the early stage (the first stage), the signs of liver toxicity Such as abdominal pain, persistent vomiting, jaundice, right upper quadrant pain, etc., will become more obvious 24 to 48 hours after a large amount of intake (the second stage). Serum transaminase usually begins to rise 16 hours after taking the drug, and clinical symptoms begin to appear. The third phase begins to appear about 3-4 days after taking the drug, at which time the degree of liver damage and prognosis can be predicted. Symptoms of liver toxicity can range from mild symptoms with elevated liver function values ​​(AST > 1,000IU / L) to severe fulminant hepatitis with metabolic acidosis, jaundice, hypoglycemia, AST > 10,000IU / L, coagulation abnormalities and hepatic encephalopathy. The fourth stage can cause oliguric renal failure, which can lead to death in severe cases.

[0005] Some patients with acetaminophen poisoning have mild liver damage but severe nephrotoxicity. The main reason is that APAP is directly metabolized by cytochrome P-450s (cytochrome P450s, C VPs) on the renal tubules to cause nephrotoxicity. However, acute renal failure may also be caused by acute liver failure, primary kidney injury (FeNa > l) or hepatorenal syndrome (FeNa < 1). The calculation formula of FeNa: (Sodiumurinary ÷ Creatinineurinary) ÷ (Sodiumplasma ÷ Creatinineplasma) χ 100.

[0006] The peak blood concentration of acetaminophen can be reached within 1 to 2 hours after oral administration, most of which are metabolized from the liver, and more than 90% are combined with glucuronide and sulfate to form non-toxic metabolites. Only less than 5% are metabolized by different CYPs, including: CYP2E1, CYP1A2 and CYP3A4, among which CYP2E1 and CYP1A2 are the main metabolic enzymes. The metabolite N-acetyl-p-benzoquinoneimine, or NAPQI (as shown in Figure 1 ) produced by the metabolism of these enzymes is a highly active electrophile. Under normal circumstances, NAPQI will immediately react with intracellular glutathione to form a non-toxic thiol compound (mercaptide). When acetaminophen is used in excess, the consumption rate of glutathione in the cell is greater than the synthesis rate, and when the content of glutathione in the cell is lower than 30% of the normal level, NAPQI will bind to macromolecules or nucleotides containing cysteine ​​in the cell, lead to liver cell damage. From the staining of cell tissue, it can be proved that before the necrosis of liver cells, NAPQI will form a covalent bond with the thiol group of cysteine ​​in the center of the liver lobule.

[0007] For patients with liver disease, alcoholism, or taking drugs that induce liver cytochrome P450 enzyme activity, ^mouth: Carbamazepine, Ethanol, Isoniazid, Phenobarbital (possibly other barbiturates), Phenytoin, Sulfinpyrazone, Sulfonylureas ^ Rifampin, Primidone, etc. All belong to the high-risk group that will cause severe hepatotoxicity of APAP. At this time, if the patient has complications such as adult respiratory distress syndrome, cerebral edema, uncontrollable hemorrhage, infection or multiple organ failure, it is easy to cause death. Taking alcohol as an example, alcohol is mainly metabolized by CYP2E1 in the liver, and its mechanism of inducing APAP poisoning can be divided into three stages. The first stage is that alcohol and APAP compete for the receptor of CYP2E1 in the liver. At this time, the concentration of NAPQI will first The second stage is that alcohol will prolong the half-life of CYP2E1 from 7 hours to 37 hours, which will increase the content of CYP2E1 in the liver, and the concentration of NAPQI will slowly rise at this time; the third stage is that when the alcohol is withdrawn, the liver has more More CYP2E1 to metabolize acetamide Acetaminophen can significantly increase the toxic metabolites of acetaminophen and cause liver cell damage. Studies in recent years have confirmed that the use of diallyl sulfide can effectively prevent hepatotoxicity caused by acetaminophen in mice, and further found that diallyl sulfide can inhibit the activity of CYP2E1 enzymes. The protective mechanism of hepatotoxicity should be through the inhibition of acetaminophen to produce the active intermediate product NAPQI.

[0008] The liver function of rats is tested by invasive and non-invasive methods to monitor the development of liver damage and screen for liver diseases. The most commonly used methods include measuring serum aspartate aminotransferase (AST), Alanine aminotransferase (ALT) and alkaline phosphatase (alkaline phosphatase) values, and the measurement of liver cell products such as: bilirubin (bilirubin), albumin (albumin), and the use of prothrombin The prothrombin time is used to detect coagulation factors, etc.; the quantitative test of liver function is based on the concentration of substances in the serum that are almost only metabolized by the liver. Hepatic blood flow is related to the quantity of substances supplied to the liver, and conversely, the clearance of the substances depends on the ability of the liver to metabolize.

[0009] Galactose is a sugar with a high extraction ratio, 90% of which is metabolized in the liver. In the liver, galactose is produced by galactokinase through epimerization reaction ( epimerization) to convert it into Glucose-1-phosphate; the action of galactokinase is the rate-limiting step of the galactose metabolic pathway in hepatocytes. The high extraction rate of galactose makes galactose metabolism, which depends on liver blood flow and liver function, the most important way to detect liver function. At present, there is no certain rule to evaluate the residual liver function of rats. Measurement The ability to metabolize a given compound (eg, galactose) allows the inference of a rate-determining step in the metabolism in the liver, and it is possible to obtain a representative quantity of residual liver function.

[0010] The inventor of this case tested patients with chronic hepatitis, liver cirrhosis and liver cancer with the galactose single-point method, and the results showed that the galactose single-point method can accurately detect these liver diseases; the galactose single-point method has been successfully applied to the exclusion of patients with liver diseases Residual liver function from drugs such as promazine and the antibiotic cefoperazone. In addition, the galactose single-point method has been recommended as one of the methods for testing liver function in the Guidance for Industry published by the US Food and Drug Administration (FDA). It can be seen that the above-mentioned customary usage of acetaminophen still has many deficiencies, is not a good design, and needs to be improved. Contents of the invention

[0011] In view of the shortcomings of side effects such as liver toxicity caused by the above-mentioned customary use of acetaminophen, the inventor of this case desperately wanted to improve and innovate, and after years of research, he finally successfully developed this new acetaminophen without hepatic side effects Compound.

[0012] The object of the present invention is to provide a new compound combination for improving hepatotoxicity of paracetamol (Acetaminophen, APAP) drug without liver side effects, comprising: (a)—acetaminophen in a pharmaceutically effective amount; And (b)—or two or more compounds that can reduce the hepatotoxicity of drugs metabolized by liver enzyme CYP2E1.

[0013] In order to achieve the purpose of the aforementioned invention, wherein the compound is at least one selected from the following groups or any combination thereof: polyoxyethylene (20) sorbitan monolaurate (Polyethylene glycol sorbitan monolaurate; Tween 20), Microcrystalline Township Microcrystalline cellulose, Dicalcium phosphate dihydrate, polyoxyethylene 23 lauryl ether (Brij 35), saccharin, mannitol, polyoxyethylene alkyl ether ( Cremophor RH40), Sucralose, Crospovidone, Sodium starch glycolate, Acrylic resin S100 (Eudragit S100), Sodium carboxymethyl cellulose (Croscarmellose sodium), Polyoxyethylene polyoxy Propylene (Pluronic F68), Menthol, Low-substituted hydroxypropyl cellulose, Pregelatinized starch, Dextrates NF hydrated, Citric acid, Castor oil poly Oxyethylene ether (Cremophor EL), colloidal silicon dioxide (Aerosil 200), polyethylene glycol monostearate (Myrj 52), sorbic acid (Sorbic acid), lemon oil (Lemon oil), hydroxypropyl cellulose Hydroxypropyl cellulose, Sorbitol, Acesulfame potassium, Hydroxypropyl methylcellulose, Lactose monohydrate, Maltodextrin , Brij 58, Brij 76, Tween 80, Tween 40, PEG 400, PEG 4000, PEG 8000, Span 60, Sodium benzoate (Sodium benzoate), Hydroxy ethylmethylcellulose phthalate, Methylcellulose phthalate, Span 80, Sodium cyclamate, Glyceryl behenate, Oxide red, Glycerin monostearate, Copovidone K28, Starch acetate, Magnesium stearate, Sodium lauryl sulfate, Providone K30, and PEG 2000.

[0014] In order to achieve the purpose of the aforementioned invention, wherein the composition of the compound is at least one selected from the following groups or any combination thereof, and the effective dosage of the composition is limited: Polyoxyethylene (20) sorbose monolaurate unitary (Polyethylene Glycol sorbitan monolaurate; Tween 20) in which the content is 0.17-5.5 grams, microcrystalline cellulose (Microcrystalline cellulose) in which the content is 100-1000 mg, dicalcium phosphate dihydrate in which the content is 10-250 mg, polyoxygen Ethylene 23 lauryl ether (Brij 35) contains 100-1000 mg, saccharin (Saccharin) contains 10-40 mg, mannitol (Mannitol) contains 10-250 mg, polyoxyethylene alkyl ether (Cremophor RH40) wherein the content is 0.17~5.5 grams, sucralose (Sucralose) wherein the content is 10~250 mg, pyrrolidone (Crospovidone) wherein the content is 0.17~5.5 grams, sodium starch glycolate (Sodium starch glycolate) wherein the content is 0.17~ 5.5 grams, acrylic resin S100 (Eudragit S100) with a content of 0.17 to 5.5 grams, carboxymethyl cellulose sodium (Croscarmellose sodium) with a content of 0.17 to 5.5 grams, polyoxyethylene polyoxypropylene (Pluronic F68) with a content of 1.4 ~5.5 grams, menthol (Menthol) containing 8~34 mg, low-substituted hydroxypropyl cellulose (Low-substituted hydroxypropyl cellulose) containing 0.19~0.82 grams, pregelatinized starch (Pregelatinized starch) containing 1.7 ~5.5 grams, Dextrates NF hydrated content is 0.17~5.5 grams, citric acid (Citric acid) content is 10~42 mg, castor oil polyoxyethylene ether (Cremophor EL) content is 1.7~5.5 grams, colloidal two Silicon oxide (Aerosil 200) with a content of 0.17~5.5 grams, polyethylene glycol monostearate (Myrj 52) with a content of 1.4~5.5 grams, sorbic acid (Sorbic acid) with a content of 6~24 mg, lemon Oil (Lemon oil) wherein the content is 0.17-5.5 grams, hydroxypropyl cellulose (Hydroxypropyl cellulose) wherein the content is 0.17-5.5 grams, sorbitol (Sorbitol) wherein the content is 0.17-5.5 grams, acesulfame potassium (Acesulfame potassium) ), the content of which is 1.4-5.5 grams, and the content of hydroxypropyl methylcellulose phthalate (Hydroxypropyl methylcellulose) is 0.17-5.5 grams. Lactose monohydrate contains 6~24 mg, Maltodextrin contains 0.17~5.5 grams, Brij 58 contains 0.17~5.5 grams, Brij 76 contains 0.17~5.5 grams, Tween 80 The content of which is 0.17~5.5 grams, the content of Tween 40 is 1.4~5.5 grams,? £0 400, whose content is 1.4-5.5 grams, PEG 4000, whose content is 1.4-5.5 grams, PEG 8000, whose content is 1.4-5.5 grams, Span 60, whose content is 1.4-5.5 grams, sodium benzoate (Sodium benzoate) The content of 2.9~11.9 mg, the content of Hydroxy ethylmethylcellulose phthalate (Hydroxy ethylmethylcellulose) is 0.17~5.5 grams, the content of Methylcellulose phthalate (Methylcellulose) is 0.17~5.5 grams, the content of Span 80 is 1.4 ~5.5 grams, Sodium cyclamate (Sodium cyclamate) containing 3.3~13.2 mg, Glyceryl behenate (Glyceryl behenate) containing 17.4~69.9 mg, Iron oxide red (Oxide red) containing 11.3-45.2 mg, Glycerin monostearate (Glycerin monostearate) contains 1.4~5.5 grams, Copovidone K28 contains 0.17~5.5 grams, Starch acetate (Starch acetate) contains 0.17~5.5 grams, Magnesium stearate (Magnesium stearate) ) with a content of 9.7-39.0 mg, sodium lauryl sulfate with a content of 4.7-19.0 mg, Providone K30 with a content of 0.18-0.73 mg, and PEG 2000 with a content of 1.4-5.5 g.

[0015] In order to achieve the purpose of the aforementioned invention, wherein the compound is selected from mannitol (Mannitol) and combined with sucralose (Sucalose) is the best.

[0016] In order to achieve the aforementioned object of the invention, wherein the combinations are used separately, simultaneously or sequentially.

[0017] In order to achieve the aforementioned object of the invention, wherein the acetaminophen and / or the compound that can reduce the hepatotoxicity produced by the drug metabolized by the liver enzyme CYP2E1 is in the form of gel, spray, pastille, lozenge or dispersible tablet given in the form of.

[0018] In order to achieve the aforementioned object of the invention, wherein the combination is contained in a medicine pack, a set or a patient pack. Another object of the present invention is to provide a use of the combination in the manufacture of a medicament for improving the hepatotoxicity of acetaminophen drugs without hepatic side effects. Description of drawings

[0019] Figure 1 is a diagram of the metabolic pathway of acetaminophen (APAP) in the liver; Figure 2 is (A) normal control group, (B) APAP hepatotoxicity group, (C) dicalcium hydrogen monophosphate, (D) mannitol, (E) menthol, (F) sucralose, (G) mannose Alcohol + sucralose (1.67 + 1.67 mg / kg) and (H) mannitol + sucralose (0.83 + 0.83 mg / kg) liver protection test group, rat liver tissue slices after tube feeding a single dose: (A ) Liver tissue type in the normal control group; (B) The liver cells in the peripheral central vein (V) were fragmented, and there were inflammatory cells, necrosis and vacuolation; compared with the APAP hepatotoxicity group, liver protection In the test group, except the dicalcium hydrogen phosphate test group, the liver cells around the central vein of the other test groups were relatively complete, with obvious nuclei and fewer vacuoles (D, E, F, G, H), and ( F), (G) are closest to normal rat liver tissue (H&E staining, 200X). detailed description

[0020] The present invention will be further described with reference to the following examples, but these examples are for illustrative purposes only, and should not be construed as limiting the implementation of the present invention.

[0021] In view of the shortcomings of side effects such as liver toxicity caused by the above-mentioned customary use of acetaminophen, the inventor of this case desperately wanted to improve and innovate, and after years of painstaking research, he finally successfully developed this acetaminophen without liver side effects Phenol new compound.

[0022] In order to achieve the above-mentioned purpose of the new paracetamol compound without hepatic side effects, the present invention first uses the new paracetamol compound to induce hepatotoxicity in rats as a model, and studies the combined use of one or any combination of CYP2E1 inhibitors The effect on the hepatotoxicity induced by acetaminophen in rats; In addition to using general hepatotoxicity markers and pathological tissue sections, the residual liver function of rats was quantitatively measured by galactose single-point method (GSP), and further Evaluate.

[0023] All technical and scientific terms described in this specification, unless otherwise defined, have meanings commonly understood by those of ordinary skill in the art.

[0024] As used herein, the term "combination (combination)", when applied to two or more compounds and / or agents (also referred to herein as ingredients), is intended to define that there are two or more compound / agent combination materials. The terms "combined" and "combining" are to be construed accordingly herein.

[0025] Pharmaceutical kits, pharmaceutical packs or patient-packs (patient packs) in which two or more compounds / agents are co-packaged or co-presented (eg, as part of a unit-dose batch).

[0026] The present invention is illustrated by the following examples, but the present invention is not limited by the following examples. The medicines and biological materials used in the present invention are commercially available and easy to obtain, and the following are only examples of available channels.

[0027] Example 1

[0028] Acetaminophen is used in combination with one or any combination of safe and pharmaceutically acceptable common excipients to improve drug hepatotoxicity in animal experiments.

[0029] Materials and Methods

[0030] 1. Test material

[0031] All organic solvents were of HPLC grade, purchased from Tedia Co., Ltd. (Fairfield, OH, USA), APAP was purchased from Sigma Chemical Company (St. Louis, MO USA), and the galactose injection solution was prepared by Nanguang Chemical Pharmaceutical Co., Ltd. Dissolve 400 grams of galactose (Sigma) in 1 liter of distilled water containing an appropriate buffer solution system and isotonic salts for injection.

[0032] 2. Try a dangerous animal

[0033] Male SD (Sprague-Dawley) rats with a body weight of 175-280 grams were purchased from the National Laboratory Animal Center (Taiwan). The animal experiments were carried out in accordance with the animal experiment guidelines of the National Institute of Health. All rats were placed in an air / humidity conditioned environment Under the condition of 12 hours of light and 12 hours of darkness each, the supply of water and feed is not limited. During the experiment, the body weight of the rats was continuously monitored, and the water was supplied as usual.

[0034] 3. Experimental Treatment

[0035] Select and screen effective inhibitors of CYP2E1 in vitro, and carry out animal experiments to improve liver toxicity caused by APAP without taking or taking inhibitors together. Hepatotoxicity test animals were given a single dose of APAP 1000 mg per kg body weight by gavage to produce hepatotoxicity. For liver protection experimental animals, rats were given 1.67 mg of dicalcium phosphate per kilogram of body weight; or 1.67 mg of mannitol per kilogram of body weight; or 1.67 mg of menthol per kilogram of body weight; or 1.67 mg of menthol per kilogram of body weight; 1.67 mg of HUEXC041 for feeding; or 1.67 mg of mannitol and 1.67 mg of sucralose per kg body weight by tube feeding; or 0.83 mg of mannitol and 0.83 mg of sucralose per kg body weight by tube feeding; or 0.42 mg mannitol and 0.42 mg sucralose per kg body weight tube-fed; or 0.17 mg mannitol and 0.17 mg sucralose per kg body weight tube-fed and a single dose of APAP 1000 mg / kg body weight tube-fed. Measure the aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the plasma to represent the liver inflammation index; measure the galactose single-point method (GSP) before and 16 hours after administration ), to analyze and quantify the remaining liver function of the rats; at the same time, analyze the pathological changes of the liver histopathology in each group of test rats, and evaluate the mechanism of liver injury or liver protection.

[0036] 4. Blood samples

[0037] After the treatment, the rat was anesthetized with acetic acid and sacrificed. The blood was drawn from the tail artery of the rat and placed in a test tube containing EDTA. The plasma (plasma) was centrifuged at 13,000g at 4°C for 15 minutes, and the separated plasma was divided into microvolumes. tubes (Eppendorf tubes) and stored at -80°C.

[0038] 5. Biochemical analysis

[0039] Liver cell damage is quantified by measuring the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in plasma. AST and ALT activities are commonly used indicators of liver toxicity. Synchron LXi 725 system to measure (Beckman Instruments, USA).

[0040] 6. Optical Microscopy

[0041] Histological analysis of the liver was carried out immediately after the sacrifice of rats; liver samples were fixed with 10% phosphate-buffered formalin, then dehydrated and embedded in paraffin at 5 μm Thickness sections were stained with hematoxylin and eosin, and the glycogen staining test (Periodic acid Schiff stain, PAS) was performed. After staining, histological observation was performed with an optical microscope.

[0042] 7. Quantitative Testing of Liver Function

[0043] After the test, all rats were subjected to the galactose single-point method (GSP) test, and the rats received a rapid intravenous injection within 30 seconds, injecting 0.4 g / ml BW galactose solution 0.5 g / kg; Blood samples were taken at 10, 15, 30, 45 and 60 minutes each, and the blood samples were taken from the tail vein; the galactose content was measured by the colorimetric galactose dehydrogenase method, and the test concentration ranged from 50 to 1,000 g / ml, the within-day variation of each concentration is calculated by the standard deviation (standard deviation) and the coefficient of variation (coefficient of variation, CV) percentage, The maximum allowable coefficient of variation is 10% CV; the day-to-day variation is tested by comparing the slope and intercept of calibration curves; the galactose single-point method (GSP) is 30 Concentration of galactose in blood at 60 minutes after cessation of second injection.

[0044] 8. Statistical analysis

[0045] All the data are expressed as mean ± standard deviation (SD), and the test results are calculated by the one-way analysis of variance (ANOVA) test method to calculate whether there is a statistically significant difference, using the Statistical Package of the Social Science program (Version 13, SPSS Inc.) software package to calculate; then use post hoc test (post hoc test) least significant difference (least significant difference) method#丧multiple comparisons to confirm the significant difference between the groups; the group mean significant difference is; ? <0.05 .

[0046] result

[0047] 1. Results of biochemical analysis

[0048] At the end of the test, the body weight and relative liver weight of the test animals were measured, and there was no significant difference compared with the normal control group animals; the blood biochemical analysis results are shown in Table 1, except for the aspartic acid in the plasma of the APAP hepatotoxicity group The activities of transaminase (AST) and alanine transaminase (ALT) were significantly higher than those in the normal control group (the AST activity in the plasma of the normal control group was 202±34 IU / L; the AST activity in the plasma of the APAP liver toxicity group was 499±112 KS / , p < 0.005; the ALT activity in the plasma of the normal control group was 56±14 IU / L; the ALT activity in the plasma of the APAP liver toxicity group was 368±71 IU / L, p < 0.005), showing that APAP The hepatotoxicity group produced biochemical liver damage; except that the dicalcium hydrogen monophosphate test group was not as expected, this liver damage phenomenon can be improved by the simultaneous use of commonly used safe excipients such as mannitol, menthol, and sucralose. The measured liver inflammation indexes AST, ALT, GSP and the total HAI-score evaluation of the severity of liver lesions on the representative pathological tissue slices all decreased significantly. The experimental data are shown in Table 1, and the combination of mannitol and sucralose Best, no different from the normal control group.

[0049] Table 1 The normal control group, the APAP hepatotoxicity group, and the hepatoprotective test groups of dicalcium hydrogen monophosphate, mannitol, menthol, sucralose, etc., after tube-feeding a single dose test, the galactose single point method (GSP) Analysis, aspartate aminotransferase (AST), alanine aminotransferase (ALT) activity and pathological tissue section Total HAI-score, the value is calculated as mean ± SD Liver function parameters GSP (mg / L) AST (IU / L) ALT (IU / L) Total

[0050] HAI-score

[0051] 1. Normal control group (n=6) 289 ±38 202 ± 34 56 ± 14 0.0 ±0.0

[0052] 2. APAP liver toxicity group (n=6) 848 ± 123 499 ± 112 368 ±71 4.9 ± 1.8

[0053] 3. Dicalcium hydrogen monophosphate (1.67

[0054] 444 ± 60*** 315 ±42* 196 ±65* 3.1± 1.1* mg / kg) test - test group (; n=6)

[0055] 4. Mannitol (1.67 mg / kg)

[0056] 253 ±29*** 201 ± 30*** 79± 34*** 0.8± 0.3*** Test group (n=6)

[0057] 5. Menthol (1.67 mg / kg)

[0058] 289 ± 20*** 187 ± 21*** 109±23*** 1.1± 1.2** Test group (n=6)

[0059] 6. Sucralose (1.67

[0060] 218 ±31*** 199 ± 24*** 83 ± 23*** 0.6 ± 0.4*** mg / kg) test - test group (; n=6)

[0061] 7. Mannitol + Sucralose

[0062] (1.67 + 1.67 mg / kg) Test group 236 ±33*** 198 ± 37*** 59± 13*** 0.5± 0.4***

[0063] (n=6)

[0064] 8. Mannitol + Sucralose

[0065] (0.83 + 0.83 mg / kg) Test group 244 ± 19*** 190 ±23*** 65 ± 19*** 0.7 ±0.5***

[0066] (n=4)

[0067] 9. Mannitol + Sucralose

[0068] (0.42 + 0.42 mg / kg) Test group 281 ± 58*** 187 ± 41*** 96 ± 14*** 1.4± 1.7*

[0069] (n=4)

[0070] 10. Mannitol + Sucralose

[0071] (0.17 + 0.17 mg / kg) Test group 371 ± 49*** 298 ± 49* 101±24*** 2.1± 1.2*

[0072] (n=4)

[0073] 2. Histopathology

[0074] The improved results were also reflected in the corresponding liver tissue. After a single oral dose of 1000 mg / kg APAP was gavaged in rats, hepatotoxicity was successfully produced in vivo. Visceral tissue sections showed that the hepatocytes around the central vein showed fragmentation, obvious vacuolization, few nuclei, and even some hepatocytes had necrosis symptoms. Compared with the normal control group, the liver cells were seriously injured (as shown in Figure 2B ); In contrast, the liver structure in the rats in the normal control group was relatively normal, and the liver cells were intact and arranged neatly without vacuoles (as shown in Figure 2A); while mannitol, menthol, sucralose The liver tissue section results of the rats in the liver protection test group showed that the liver cells were relatively complete, with obvious nuclei and fewer vacuoles (as shown in Figure 2C, D, F, G, H), showing mannitol, menthol, trichloro The liver tissue of rats in the sucrose and other liver protection test group was closer to the liver tissue of normal rats, and the combination of mannitol and sucralose had the best protective effect, and it was positively correlated with the dose. The higher the dose, the better the protective effect.

[0075] 3. Measurement of Residual Liver Function

[0076] As shown in Table 1, the galactose single-point method (GSP) value of rats in the normal control group and the APAP liver toxicity group had a highly significant difference (the GSP value of the normal control group rats was 289±38 mg / L; The GSP value of the rats in the toxicity group was 848±123 mg / L,;? < 0.005), in addition, the GSP values ​​of the rats in the liver protection test groups such as dicalcium hydrogenate, mannitol, menthol and sucralose were respectively 444±60 mg / L, 253±29 mg / L, 289±20 mg / L, 218±31 mg / L, compared with APAP liver toxicity group, mannitol, menthol and sucralose and other liver protection test group The GSP values ​​of the rats were significantly different from those of the APAP hepatotoxicity group rats (;? < 0.005); the GSP values ​​of the hepatotoxic rats were significantly increased when APAP was administered alone; however, mannitol, menthol and Rats in the liver protection test group with excipients such as sucralose can resist this change.

[0077] Embodiment two

[0078] Screening for inhibitors of cytochrome P450 2E1 (CYP2E1) - rat liver microsomes versus human liver microsomes. Materials and Methods

[0079] 1. Test material

[0080] In this example, microsomes prepared from the livers of rats and human species were used for in vitro screening of CYP2E1 inhibitors. Cytochrome P450 2E1 (CYP2E1) inhibitors were screened for 55 common and safe edible excipients, and the large An effective inhibitor of CYP2E1 in mouse or human liver; the screening principle of the CYP2E1 inhibitor is: use the reaction of CYP2E1 in microsomes prepared from the liver of the species with its specific substrate Chlorzoxazone (CZX), add the test sample to act, and then detect CYP2E1 Metabolite Standard 6-OH-CZX ( 6 _Hydroxy-Chlorzoxazone), and based on the 6-OH-CZX production of the control group (control), the CYP 2E1 inhibition rate of the test sample was calculated.

[0081] Each test sample was dissolved in 10% methanol (methanol) or secondary water to test the effect of different concentrations of excipients (66uM, 33uM, 16.5uM; 0.167%, 0.08%, 0.042%, w / v) on CYP2E1 Inhibition rate, the results of the tested excipients are listed in Table 2.

[0082] In this example, the agents required for screening cytochrome CYP2E1 inhibitors using rat liver or human liver microsomes are as follows:

[0083] (1) CYP2E1: 100 mM potassium phosphate ( H 7.4) contains 10 mg / mL P450 protein concentration.

[0084] (2) Control Protein: 10 mg / mL P450 Protein dissolved in 100 mM Potassium Phosphate (pH 7.4).

[0085] (3) Buffer Solution: 0.5 M Potassium Phosphate (pH 7.4).

[0086] (4) Stop Solution: ice-acetonitrile.

[0087] (5) Cofactors: Contains 100 mM NADP+ and 10 mM Glucose 6-Phosphate.

[0088] (6) Glucose 6-Phosphate Dehydrogenase: 2000 units / ml dissolved in sterile water.

[0089] (7) Chlorzoxazone: Substrate, 16 mM Chlorzoxazone dissolved in 10% methanol.

[0090] (8) DDTC (Diethyldithiocarbamic acid): CYP2E1 selective inhibitor (positive control group), 20 mM DDTC dissolved in 10% methanol (methanol).

[0091] (9) NADPH-regenerating System: Add 530 uL Cofactors, 40 uL G6PDH (Glucose 6-Phosphate Dehydrogenase Solution) and 100 uL Control Protein to 3.42 mL.

[0092] 2. Screening of Cytochrome P450 2E1 (CYP2E1) Inhibitors

[0093] The protocol for screening cytochrome CYP2E1 inhibitors using rat liver or human liver microsomes is as follows: (1) 0.1M phosphate buffer (pH 7.4) containing 0.5 mg / mL rat liver or human liver microsomes, 5 mM MgCl in a water bath at 4°C 2 Let stand for 15 minutes;

[0094] (2) At this time, the experimental group was added with cytochrome P4 5 0 2E1 Reaction matrix drug 16 mM Chlorzoxazone and the compound to be screened; the control group was replaced by methanol: sterile water = 1: 1; the positive control group was replaced by DDTC;

[0095] (3) Finally, add coenzyme 1 mM NADP+, 10 mM G6P and 2 IU G6PD. The reaction solution was transferred to a 37°C water bath for pre-incubation for 1 minute, and the reaction time for the activity test experiment was 30 minutes;

[0096] (4) After the reaction, stop the reaction with 500 acetonitrile, let the sample stand for 1 minute, add the internal standard (5 ug / mL 4-hydroxy-tobutamide), centrifuge and take 20 uL of the supernatant, and use methanol: sterile water as a For ten-fold dilution, take 5 uL of the returned solution and inject it into the LC / MS / MS system for analysis;

[0097] (5) Result analysis: After converting the signal value measured by LC / MS / MS into the production amount (pmol) of CYP2E1 metabolite standard 6-Hydroxy-Chlorzoxazone, the CYP 2E1 inhibition rate of the control group is based on the control group is 0%, the CYP 2E1 inhibition rate of each positive control group and test group is calculated with the following formula: Experimental group 6 «OH« CZ3 production

[0098] CYP2I1 inhibition rate (%) [ 1 - ,,,,,,; j 100%

[0099] Control group 6-OH- ί formation results

[0100] 1. The CYP 2E1 inhibition rate measured by the positive control group (DDTC) is shown in Table 2. It can be seen from Table 2 that when the concentration of DDTC is 100 μM, the CYP 2E1 inhibition rate can reach 89.2%. .

[0101] 2. CYP 2E1 inhibition rate of test group

[0102] The CYP 2E1 inhibition rate measured by the excipients in rat liver microsomes is shown in Table 2. From the results, it can be seen that the excipients at different concentrations (66μΜ, 33μΜ, 16.5μΜ; 0.167%, 0.08%, 0.042%, w Under the conditions of / v), it has different degrees of inhibitory effect on cytochrome P450 2E1, wherein 0.167% Brij 58 Best suppression effect (100.0±0.00%)

[0103] Table 2. Inhibition rate of excipients in rat liver microsomes for in vitro screening of CYP 2E1 inhibitors Excipient CYP2E1 inhibition rate (%)

[0104] Test Concentration 66 μM 33 μM 16.5 μM Control 0

[0105] (100 μΜ) (50 μΜ) (10 μΜ) positive control (DDTC)

[0106] 89.2±2.2 50.4±1.1 8.6±1.1

[0107] 100.0 ±0.0 98.6 ±0.2 96.5 ±0.3

[0108] Brij 58

[0109] (0.167%) (0.084%) (0.042%)

[0110] 100.0 ±0.0 98.5 ±0.1 97.5 ±0.3

[0111] Brij 76

[0112] (0.167%) (0.084%) (0.042%)

[0113] 95.9 ±0.4 92.5 ±0.7 80.6 ±1.9

[0114] Tween 20

[0115] (0.167%) (0.084%) (0.042%)

[0116] 85.1±0.4 79.4±1.4 68.2±1.3

[0117] Tween 80

[0118] (0.167%) (0.084%) (0.042%) Microcrystalline Cellulose

[0119] 78.7 ±2.8 75.0 ± 5.2 73.9 ± 1.8 Microcrystalline

[0120] (0.025%) (0.013%) (0.006%)

[0121] 78.1±1.4 71.5±0.5 54.0±3.2

[0122] Tween 40

[0123] (0.167%) (0.084%) (0.042%) Dicalcium phosphate

[0124] Dicalcium phosphate 76.7 ±0.8 62.0 ±2.6 55.0 ±4.6 dihydrate

[0125] saccharin

[0126] 67.4 ±3.9 59.6 ±3.3 35.3 ±2.0 Saccharin

[0127] 67.2±1.4 59.3±2.5 45.9±2.6

[0128] Brij 35

[0129] (0.025%) (0.013%) (0.006%) Mannitol

[0130] 60.6 ±3.3 51.0±2.7 40.9 ±2.8 Mannitol

[0131] polyoxyethylene alkyl ether

[0132] 57.4 ±3.2 49.4 ±2.9 48.0 ±2.1 Cremophor RH40 Excipient CYP2E1 inhibition rate (%)

[0133] Test Concentration 66 μM 33 μM 16.5 μM Sucralose

[0134] 54.0 ±4.2 46.8 ±0.8 41.1 ±2.7

[0135] Sucralose

[0136] 52.5 ±4.6 43.4 ±3.0 35.1 ±2.2

[0137] PEG400

[0138] (0.167%) (0.084%) (0.042%) pyridone

[0139] 48.7 ±0.4 43.2 ±3.6 41.1 ±2.7 Crospovidone

[0140] 48.1 ±2.4 39.4 ±1.8 32.7 ±0.8

[0141] PEG4000

[0142] (0.167%) (0.084%) (0.042%) Sodium starch glycolate 41.2 ±4.9 37.6 ±2.5 34.1 ±0.8 Sodium starch glycolate (0.167%) (0.084%) (0.042%) Acrylic resin SI 00 39.7 ±4.9 33.5 ±4.0 12.7 ± 1.9 Eudragit SI 00 (0.167%) (0.084%) (0.042%) Polyoxyethylene 37.3 ± 3.0 18.9 ± 1.4 14.9 ± 0.9 Pluronic F68 (0.167%) (0.084%) (0.024%) Menthol

[0143] 36.4 ±0.3 15.3 ±7.9 7.2 ±2.9 Menthol

[0144] low substituted hydrocarbon propyl cellulose

[0145] 36.2 ±6.0 33.8 ±1.4 28.7 ±2.2 Low- substituted

[0146] (0.025%) (0.013%) (0.006%) hydroxypropyl cellulose

[0147] Pregelatinized starch 33.6 ±2.0 26.2 ±2.8 14.0 ±2.5 Pregelatinized starch (0.167%) (0.084%) (0.024%)

[0148] 32.9 ±2.0 27.0 ±3.0 13.2 ±0.6

[0149] Dextrates, NF hydrate

[0150] (0.167%) (0.084%) (0.024%)

[0151] 27.6 ±3.6 12.4 ±2.2 7.5 ±2.3

[0152] Citric acid

[0153] Castor Oil Ethoxylates 25.2 ±2.7 12.9 ±2.2 5.9 ±0.3 Cremophor EL (0.167%) (0.084%) (0.024%) Excipient CYP2E1 inhibition rate (%) Test concentration 66 μΜ 33 μΜ 16.5 μΜ Colloidal silicon dioxide 23.8 ± 2.4 22.8 ± 1.7 4.7 ± 1.2

[0154] Aerosil 200 (0.167%) (0.084%) (0.024%) Polyethylene glycol monostearate 20.5 ±0.3 18.5 ±0.6 17.5 ± 1.5 Myrj 52 (0.167%) (0.084%) (0.024%)

[0155] 19.2 ±2.0 15.1 ±0.6 9.9 ±0.3

[0156] PEG 8000

[0157] (0.167%) (0.084%) (0.024%) Sorbic acid

[0158] 19.0 ± 5.6 13.2 ±4.4 12.1 ± 5.7 Sorbic acid

[0159] 4 Lemon oil 18.2 ±3.7 13.9 ±2.9 9.7 ±3.8 Lemon oil (0.167%) (0.084%) (0.024%) Hydroxypropyl cellulose 18.0 ±2.2 12.7 ± 1.9 6.7 ±0.7 Hydroxypropyl cellulose (0.167%) (0.084%) ) (0.024%)

[0160] 17.1±0.8 15.2±2.1 15.1±1.4

[0161] Span 60

[0162] (0.167%) (0.084%) (0.024%) Sorbitol 16.1 ±0.7 5.6 ±0.5 6.4 ±0.5 Sorbitol (0.167%) (0.084%) (0.024%) Sodium Benzoate

[0163] 15.8 ±0.9 7.8 ±4.1 7.1 ±2.0 Sodium benzoate

[0164] Acesulfame Potassium

[0165] 14.5 ± 1.9 7.1 ±2.3 3.9 ±2.7 Acesulfame potassium

[0166] Hypromellose Phthalate

[0167] 13.9 ±2.2 13.6 ±2.6 6.7 ±0.7 Hydroxypropyl

[0168] (0.167%) (0.084%) (0.024%) methylcellulose

[0169] Hydroxymethyl Cellulose Phthalate

[0170] 11.6±0.9 13.2±0.6 5.6±0.5

[0171] Hydroxy

[0172] (0.167%) (0.084%) (0.024%) ethylmethylcellulose

[0173] Methylcellulose phthalate 10.3 ± 1.7 5.2 ±0.3 5.0 ± 1.1 Methylcellulose (0.167%) (0.084%) (0.024%)

[0174] 9.4±0.6 8.5±1.3 10.6±1.9

[0175] Span 80

[0176] (0.167%) (0.084%) (0.024%) Excipient CYP2E1 inhibition rate (%)

[0177] Test concentration 66 μΜ 33 μΜ 16.5 μΜ cyclamate

[0178] 9.1 ±2.6 5.7 ±4.7 9.4 ±2.7

[0179] Sodium cyclamate

[0180] lactose monohydrate

[0181] 8.7 ±3.8 7.8 ±2.2 3.9 ±2.3 Lactose monohydrate

[0182] Maltodextrin 8.5 ±2.8 5.9±2.1 9.7 ± 5.6 Maltodextrin (0.167%) (0.084%) (0.024%) Glyceryl behenate

[0183] 8.2 ±2.0 3.1 ±2.5 3.1 ±0.2 Glyceryl behenate

[0184] Iron Oxide Red

[0185] 8.5 ± 5.1 10.7 ±4.1 10.3 ±2.1 Oxide red

[0186] Glycerin monostearate 6.9 ± 1.8 7.4 ±2.9 8.3 ± 5.7 Glycerin monostearate (0.167%) (0.084%) (0.024%)

[0187] 6.1 ±0.7 4.5 ±0.5 4.3 ±0.2

[0188] Copovidone K28

[0189] (0.167%) (0.084%) (0.024%)

[0190] / plant day 5.3 ±0.7 4.9 ± 1.2 5.5 ± 1.2

[0191] Starch acetate (0.167%) (0.084%) (0.024%) Magnesium stearate

[0192] 5.0 ± 1.6 3.0 ±0.7 2.0 ± 1.0 Magnesium stearate

[0193] sodium lauryl sulfate

[0194] 4.8 ± 1.2 6.4 ±0.9 4.6 ± 1.1 Sodium lauryl sulfate

[0195] Providone K30 3.2 ±0.2 2.2 ±0.1 4.7 ± 1.0 Benzyl alcohol -10.3 ±6.3 6.7 ± 1.0 8.2 ±2.0 Benzyl alcohol (0.167%) (0.084%) (0.024%) Sodium methylparaben

[0196] -21.5 ±2.0 -14.6 ±4.1 4.6 ±3.2 Methylparaben

[0197] Propylparaben

[0198] -27.3 ±3.7 -17.2 ±2.4 -4.1 ± 1.2 Propylparaben

[0199] -35.5 ±4.3 -21.0 ±4.8 -9.3 ±0.8

[0200] Solutol H15

[0201] (0.167%) (0.084%) (0.042%) Excipient CYP2E1 inhibition rate (%)

[0202] Test concentration 66 μΜ 33 μΜ 16.5 μΜ

[0203] Butylated hydroxyanisole

[0204] -85.5 ±3.9 -47.1 ±5.3 -16.8 ±2.5

[0205] Butylated hydroxylanisol

[0206] The CYP 2E1 inhibition rates of the excipients in human liver microsomes are shown in Table 3. From the results, it can be seen that the excipients at different concentrations (66 μΜ, 33 μΜ, 16.5 μΜ; 0.167%, 0.08%, 0.042%, w Under the conditions of / v), it has different degrees of inhibitory effect on cytochrome P4502E1, among which 0.167% Brij 58 has the best inhibitory effect (91.2±1.3%).

[0207] Table 3. Inhibition rate of excipients for screening CYP 2E1 inhibitors in human liver microsomes in vitro

[0208] Excipient CYP2E1 inhibition rate (%)

[0209] Test concentration 66 μΜ 33 μΜ 16.5 μΜ

[0210] Control group 0

[0211] (100 μΜ) (50 μΜ) (10 μΜ) positive control (DDTC)

[0212] 89.6±0.9 49.8±2.9 7.3±1.0

[0213] 91.2±1.3 80.5±1.1 62.6±2.1

[0214] Brij 58

[0215] (0.167%) (0.084%) (0.042%)

[0216] 86.2±1.3 75.7±1.6 69.0±3.8

[0217] Brij 76

[0218] (0.167%) (0.084%) (0.042%) Saccharin

[0219] 78.5±2.1 51.2±0.9 29.4±2.7

[0220] Saccharin

[0221] 77.3 ± 1.0 73.0 ± 1.7 42.4 ± 1.8

[0222] Brij 35

[0223] (0.025%) (0.013%) (0.006%)

[0224] 75.4±3.6 70.4±0.9 55.4±1.9

[0225] Tween 20

[0226] (0.167%) (0.084%) (0.042%)

[0227] 64.2 ± 1.5 54.8 ± 3.5 26.4 ± 1.8

[0228] PEG400

[0229] (0.167%) (0.084%) (0.042%) Microcrystalline Cellulose

[0230] 60.2 ±4.1 54.4 ±3.8 48.8 ±0.2

[0231] Microcrystalline

[0232] (0.025%) (0.013%) (0.006%) Excipient CYP2E1 inhibition rate (%)

[0233] Test concentration 66 μM 33 μM 16.5 μM Dicalcium phosphate

[0234] Dicalcium phosphate 60.1 ±0.3 56.8 ±2.2 31.2±2.9 dihydrat

[0235] Sucralose

[0236] 55.8 ±2.0 45.8 ±4.0 37.1 ±2.8 Sucralose

[0237] Mannitol

[0238] 54.5±4.2 51.2±2.1 44.8±1.8 Mannitol

[0239] polyoxyethylene alkyl ether

[0240] 50.4 ± 1.1 43.2 ± 3.1 30.2 ± 2.8 Cremophor RH40

[0241] Sodium starch glycolate 49.1 ±2.9 31.4± 5.2 38.9 ± 1.3 Sodium starch glycolate (0.167%) (0.084%) (0.042%)

[0242] 47.5±1.5 41.4±1.6 22.3±1.9

[0243] PEG2000

[0244] (0.167%) (0.084%) (0.042%)

[0245] 47.1±0.9 23.9±2.9 8.7±1.8

[0246] PEG4000

[0247] (0.167%) (0.084%) (0.042%)

[0248] 46.3±3.1 33.4±2.7 16.9±1.2

[0249] Tween 40

[0250] (0.167%) (0.084%) (0.042%) Pyridine 44.1 ±0.9 40.3 ±3.3 35.6± 1.8 Crospovidone (0.167%) (0.084%) (0.042%)

[0251] 39.1±2.4 40.6±3.8 29.0±1.7

[0252] Tween 80

[0253] (0.167%) (0.084%) (0.042%) Acrylic resin SI 00 38.3 ±0.1 35.6 ±2.4 23.2 ±3.5 Eudragit SI 00 (0.167%) (0.084%) (0.042%) Sodium carboxymethylcellulose 35.4 ±4.8 30.3 ± 5.4 8.1 ±2.3 Croscarmellose sodium (0.025%) (0.013%) (0.006%) Polyoxyethylene polyoxyethylene 31.5 ± 1.6 17.4 ±4.2 7.9 ±0.8 Pluronic F68 (0.025%) (0.013%) (0.006%) Peppermint Alcohol 30.8 ±0.3 20.8 ±2.1 10.5 ±0.4 Menthol Excipient CYP2E1 inhibition rate (%)

[0254] Test concentration 66 μΜ 33 μΜ 16.5 μΜ

[0255] Low-substituted hydrocarbon propylene cellulose 22.1 ±3.7 20.3 ± 1.8 17.5 ±2.9

[0256] Low-substituted (0.025%) (0.013%) (0.006%) hydroxypropyl cellulose

[0257] 21.1 ±4.4 14.2 ±3.6 9.4 ±0.2

[0258] PEG 8000

[0259] (0.167%) (0.084%) (0.024%) Citric acid 20.5 ± 1.8 15.5 ± 1.5 9.9±3.1

[0260] Citric acid

[0261] Castor oil ethoxylates 19.2 ±0.5 15.1 ±2.2 8.1 ±0.6

[0262] Cremophor EL (0.167%) (0.084%) (0.024%)

[0263] 19.2±1.1 14.4±3.2 12.9±0.6

[0264] Dextrates, F hydrate

[0265] (0.167%) (0.084%) (0.024%) Pregelatinized starch 18.3 ±2.4 12.8 ±0.8 9.9 ±0.1

[0266] Pregelatinized starch (0.167%) (0.084%) (0.024%) Polyethylene glycol monostearate 18.1 ±2.6 15.7 ±2.8 14.6 ± 1.7

[0267] Myrj 52 (0.167%) (0.084%) (0.024%)

[0268] 17.4 ±0.9 13.9 ±0.7 12.4 ±2.3

[0269] Span 60

[0270] (0.167%) (0.084%) (0.024%) Colloidal silica 15.7±3.4 17.8±2.1 7.8 ±0.4

[0271] Aerosil 200 (0.167%) (0.084%) (0.024%)

[0272] Sorbic acid 14.8 ±0.1 10.9 ±2.7 8.4 ± 1.6

[0273] Sorbic acid

[0274] 10.1 ±2.1 5.7 ±4.7 9.4 ±2.7

[0275] Span 80

[0276] (0.167%) (0.084%) (0.024%)

[0277] 4Ning sample oil 7.8 ±0.3 9.8 ±0.4 8.8 ± 1.1

[0278] Lemon oil (0.167%) (0.084%) (0.024%)

[0279] For a new compound combination without hepatic side effects to improve the hepatotoxicity of Acetaminophen (APAP), the effective dosage ranges of each excipient under different concentrations (66 μM, 33 μM, 16.5 μM) are respectively : Polyoxyethylene (20) sorbitan monolaurate (Polyethylene glycol sorbitan monolaurate; Tween 20) containing 0.17~5.5 grams, microcrystalline cellulose (Microcrystalline cellulose) with a content of 100~1000 mg, Dicalcium phosphate dihydrate with a content of 10~250 mg, polyoxyethylene 23 lauryl ether (Brij 35) with a content of 100~1000 mg, saccharin (Saccharin) whose content is 10~40 mg, mannitol (Mannitol) whose content is 10~250 mg, polyoxyethylene alkyl ether (Cremophor RH40) whose content is 0.17~5.5 g, sucralose (Sucralose) whose content 10-250 mg, 0.17-5.5 g of Crospovidone, 0.17-5.5 g of Sodium starch glycolate, 0.17-5.5 g of Eudragit S100, Carboxymethylcellulose sodium (Croscarmellose sodium) has a content of 0.17-5.5 grams, polyoxyethylene polyoxypropylene (Pluronic F68) has a content of 1.4-5.5 grams, menthol (Menthol) has a content of 8-34 mg, low Low-substituted hydroxypropyl cellulose (Low-substituted hydroxypropyl cellulose) with a content of 0.19-0.82 grams, pregelatinized starch (Pregelatinized starch) with a content of 1.7-5.5 grams, Dextrates NF hydrated with a content of 0.17-5.5 grams, citric acid ( Citric acid) in which the content is 10-42 mg, castor oil polyoxyethylene ether (Cremophor EL) in which the content is 1.7-5.5 grams, colloidal silicon dioxide (Aerosil 200) in which the content is 0.17-5.5 grams, polyethylene glycol Monostearate (Myrj 52) contains 1.4-5.5 grams, sorbic acid contains 6-24 mg, lemon oil contains 0.17-5.5 grams, hydroxypropyl cellulose (Hydroxypropyl cellulose) wherein the content is 0.17~5.5 grams, sorbitol (Sorbitol) wherein the content is 0.17~5.5 grams, acesulfame potassium (Acesulfame potassium) wherein the content is 1.4~5.5 grams, hydroxy Propyl methylcellulose phthalate (Hydroxypropyl methylcellulose) contains 0.17-5.5 grams, Lactose monohydrate (Lactose monohydrate) contains 6-24 mg, maltodextrin (Maltodextrin) contains 0.17-5.5 grams, Brij 58 The content of Brij 76 is 0.17-5.5 grams, the content of Brij 76 is 0.17-5.5 grams, the content of Tween 80 is 0.17-5.5 grams, the content of Tween 40 is 1.4-5.5 grams, the content of PEG 400 is 1.4-5.5 grams, PEG 4000 The content of PEG 8000 is 1.4-5.5 grams, the content of PEG 8000 is 1.4-5.5 grams, the content of Span 60 is 1.4-5.5 grams, sodium benzoate (Sodium benzoate) is 2.9-11.9 mg, hydroxyethyl cellulose Hydroxy ethylmethylcellulose with a content of 0.17-5.5 grams, Methylcellulose with a content of 0.17-5.5 grams, Span 80 with a content of 1.4-5.5 grams, Sodium cyclamate with a content of 0.17-5.5 grams 3.3~13.2mg, Glyceryl behenate (Glyceryl behenate) contains 17.4~69.9mg, Iron oxide red (Oxide red) contains 11.3~45.2mg, Glycerin monostearate (Glycerin monostearate) contains 1.4~5.5g, Copovidone K28 containing 0.17~5.5g, Starch acetate containing 0.17~5.5g, Magnesium stearate containing 9.7~39.0mg, sodium lauryl sulfate (Sodium lauryl sulfate), the content of which is 4.7-19.0 mg, the content of Providone K30, which is 0.18-0.73 mg, and the content of PEG 2000, which is 1.4-5.5 grams.

[0280] When the new paracetamol compound formula without hepatic side effects provided by the present invention is compared with the test results of using paracetamol alone, the biochemical analysis (ALT, AST value), pathological analysis and residual liver function The analysis results of measurement (GSP value) and other aspects all have the effect of significantly reducing the side effects of liver toxicity caused by the use of acetaminophen.

[0281] The above detailed description is a specific description of a feasible embodiment of the present invention, but this embodiment is not intended to limit the protection scope of the present invention, any equivalent implementation or change that does not depart from the technical spirit of the present invention, for example: Acetaminophen, cytochrome P450 2E1 inhibitors, the types of cytochrome P450 2E1 inhibitors selected, the concentration and ratio of application, and the equivalence examples of other changes should all be included in the scope of protection of this case.

[0282] To sum up, this case is not only innovative in the application of acetaminophen, but also uses common and safe common excipients to improve the hepatotoxic side effects caused by the use of acetaminophen.

Claims

Claims 1. A novel hepatotoxic combination therapy for improving hepatotoxicity of acetaminophen (APAP) without hepatic side effects, comprising: (a) — A pharmaceutically effective amount of p-acetaminophen; and (b) — or two or more compounds that can reduce the hepatotoxicity of drugs metabolized by the liver enzyme CYP2E1.

2. The novel compound formulation for improving hepatotoxicity of acetaminophen as described in claim 1, wherein the compound is selected from at least one or any combination thereof from the group consisting of: polyethylene glycol sorbitan monolaurate (Tween 20), microcrystalline cellulose, dicalcium phosphate dihydrate, polyethylene 23 lauryl ether (Brij 35), saccharin, mannitol, polyethylene alkyl ether (Cremophor RH40), sucralose, pyrrolidone, sodium starch glycolate, eudragit S100, sodium carboxymethyl cellulose (Croscarmellose), and polyethylene polyoxypropylene (Pluronic F68). Menthol, low-substituted hydroxypropyl cellulosic acid, pregelatinized starch, dextrates NF hydrated, citric acid, Cremophor EL, Aerosil 200, Myrj 52, sorbic acid, lemon oil, hydroxypropyl cellulose, sorbitol, acesulfame potassium, hydroxypropyl methylcellulose, lactose monohydrate, maltodextrin, Brij 58, Brij 76,Tween 80, Tween 40, PEG 400, PEG 4000, PEG 8000, Span 60, Sodium benzoate, Hydroxyethylmethylcellulose, Methylcellulose, Span 80, Sodium cyclamate, Glyceryl behenate, Oxide red, Glycerin monostearate, Copovidone K28, Starch acetate, Magnesium stearate, Sodium lauryl sulfate, Providone K30, and PEG 2000.

3. The novel hepatotoxicity-free compound combination for improving the hepatotoxicity of acetaminophen (APAP) as described in claim 2, wherein the compound composition is selected from at least one or any combination of the following groups, and the effective dosage of the composition is defined as follows: Polyethylene glycol sorbitan monolaurate (Tween 20) in a content of 0.17-5.5 g, Microcrystalline cellulose in a content of 100-1000 mg, Dicalcium phosphate dihydrate in a content of 10-250 mg, Polyethylene 23 lauryl ether (Brij 35) in a content of 100-1000 mg, Saccharin in a content of 10-40 mg, Mannitol in a content of 10-250 mg, Polyethylene alkyl ether (Cremophor) The following components are listed: RH40 (0.17-5.5 g), sucralose (10-250 mg), crospovidone (0.17-5.5 g), sodium starch glycolate (0.17-5.5 g), Eudragit S100 (0.17-5.5 g), croscarmellose sodium (0.17-5.5 g), Pluronic F68 (1.4-5.5 g), menthol (8-34 mg), low-substituted hydroxypropyl cellulosic acid (0.19-0.82 g), and pregelatinized starch (1.7 g). 5.5g, Dextrates NF hydrated (content 0.17~5.5g), Citric acid (content 10~42mg), Cremophor EL (content 1.7~5.5g), and Aerosil 200 (content 0.17~5g).5g, polyethylene glycol monostearate (Myrj 52) with a content of 1.4-5.5g, sorbic acid with a content of 6-24mg, lemon oil with a content of 0.17-5.5g, hydroxypropyl cellulose with a content of 0.17-5.5g, sorbitol with a content of 0.17-5.5g, acesulfame potassium with a content of 1.4-5.5g, hydroxypropyl methylcellulose with a content of 0.17-5.5g, lactose monohydrate with a content of 6-24mg, maltodextrin with a content of 0.17-5.5g, Brij 58 contains 0.17-5.5 grams; Brij 76 contains 0.17-5.5 grams; Tween 80 contains 0.17-5.5 grams; Tween 40 contains 1.4-5.5 grams; PEG 400 contains 1.4-5.5 grams; PEG 4000 contains 1.4-5.5 grams; PEG 8000 contains 1.4-5.5 grams; Span 60 contains 1.4-5.5 grams; Sodium benzoate contains 2.9-11.9 mg; Hydroxyethylmethylcellulose phthalate contains 0.17-5.5 grams; Methylcellulose phthalate contains 0.17-5.5 grams; Span The following compounds contain 1.4–5.5 g of the following: sodium cyclamate (3.3–13.2 mg), glyceryl behenate (17.4–69.9 mg), oxide red (11.3–45.2 mg), glycerin monostearate (1.4–5.5 g), copovidone K28 (0.17–5.5 g), starch acetate (0.17–5.5 g), and magnesium stearate (9.7–39 g).0 mg, sodium lauryl sulfate (4.7-19.0 mg), Providone K30 (0.18-0.73 mg), and PEG 2000 (1.4-5.5 g).

4. The combination of compounds as described in claim 2 for improving hepatotoxicity of acetaminophen (APAP) without hepatic side effects, wherein the compounds are preferably selected from mannitol and combined with sucralose.

5. The novel hepatotoxic combination of acetaminophen as described in claim 1, wherein the combination is used separately, simultaneously or sequentially.

6. The novel hepatotoxic combination for improving hepatotoxicity of acetaminophen as described in claim 1, wherein the acetaminophen and / or the compound that reduces hepatotoxicity caused by drugs metabolized by the liver enzyme CYP2E1 is administered in the form of gel, spray, soft lozenge, tablet or dispersible tablet.

7. The novel hepatotoxic combination of claim 1 for improving hepatotoxicity of acetaminophen, wherein the combination is contained in a medicine package, kit, or patient pack.

8. Use of the combination as described in claim 1 in the manufacture of a medicament for treating hepatotoxicity of acetaminophen drugs without hepatotoxic side effects.