CN117442575A

CN117442575A - Vanilla pharmaceutical composition and preparation and application thereof

Info

Publication number: CN117442575A
Application number: CN202311558309.8A
Authority: CN
Inventors: 魏彦君; 巴萨瓦拉杰·希达贡纳瓦; 孙中亚; 裴慧; 刘希望; 邢艳平; 徐青景; 刘艳; 胡青燕
Original assignee: Shandong Viwit Baike Pharmaceutical Co ltd; Weizhi Pharmaceutical Co ltd; Shandong Weizhi Zhongke Pharmaceutical Co ltd
Current assignee: Shandong Viwit Baike Pharmaceutical Co ltd; Weizhi Pharmaceutical Co ltd; Shandong Weizhi Zhongke Pharmaceutical Co ltd
Priority date: 2021-08-27
Filing date: 2021-08-27
Publication date: 2024-01-26
Also published as: CN113908131A

Abstract

The invention discloses a valicarb isopropyl pharmaceutical composition, and preparation and application thereof. According to the invention, the content of nitrosamine impurities in bulk drugs is controlled, and pharmaceutically acceptable acid is added into a pharmaceutical composition of a secondary amine compound (valicarb or pharmaceutically acceptable salt thereof), so that the generation of nitrosamine impurities in the preparation process and/or the storage process is effectively inhibited and reduced, the stability of the pharmaceutical composition of the secondary amine compound is improved, and the content of nitrosamine toxic impurities is controlled at a lower level, so that the pharmaceutical composition of the secondary amine compound (valicarb or pharmaceutically acceptable salt thereof) with low content of nitrosamine impurities is obtained, and the safety requirements and the regulatory requirements (for example, the limit requirements of FDA) of pharmaceutical products are met, so that the medication safety of patients can be better ensured.

Description

Vanilla pharmaceutical composition and preparation and application thereof

The application is a divisional application of an invention patent of a valicarb pharmaceutical composition capable of reducing nitrosamine impurity generation, and preparation and application, wherein the application is applied for the application of the invention of day 2021, day 08 and day 27, application number 202110997332.1.

Technical Field

The invention belongs to the field of medicines, and particularly relates to a valicarb isopropyl pharmaceutical composition, and preparation and application thereof.

Background

Valicarb is an aryl fused azapolycyclic compound capable of modulating cholinergic activity and sold as tartrate, commercially available under the name or capable of binding to neuronal nicotinic acetylcholine specific receptor sites, useful in the following diseases:

treatment of inflammatory bowel diseases (including but not limited to: ulcerative colitis, pyoderma gangrenosum and Crohn's disease), irritable bowel syndrome, spasmodic dystonia, chronic pain, acute pain, diarrhea (celiac serum), cystitis (pouttices), vasoconstriction, anxiety, panic disorder, depression, bipolar disorder, autism, sleep disorder, jet lag, amyotrophic Lateral Sclerosis (ALS), cognitive dysfunction, drug/toxin-induced cognitive impairment (e.g., due to alcohol, barbiturates, vitamin deficiency, recreational drugs (recreational drug), lead, arsenic, mercury), disease-induced cognitive impairment (e.g., caused by Alzheimer's disease (Alzheimer's disease), vascular dementia (vascular dementia), parkinson's disease, multiple sclerosis, AIDS, (large) encephalitis, trauma, renal and hepatic encephalopathy, hypothyroidism, pick's disease, korscheff's syndrome and forehead (front) and subcortical dementia (subcortical dementia)), hypertension, bulimia, anorexia, obesity, arrhythmia, gastric acid hypersecretion, ulcers, pheochromocytoma, progressive supranuclear palsy (progressive supramuscular palsy), chemical dependence and addiction (e.g., to nicotine (and/or tobacco products), alcohol, benzodiazepines, barbiturates, opioids or cocaine dependence and addiction), headache, migraine, stroke, traumatic Brain Injury (TBI), obsessive-compulsive disorder (OCD), psychosis, huntington's chorea, tardive dyskinesia, hyperkinesias, dyslexia, schizophrenia, multiple sclerosis dementia, age-related cognitive decline, epilepsy, including seizure deficiency (including petit mal absence epilepsy), attention deficit hyperactivity disorder (attention deficit hyperactivity disorder) (ADHD), tourette's Syndrome (Syndrome); in particular, nicotine dependence, addiction and withdrawal, including use in smoking cessation therapies (see: CN1509174 a).

Nitrosamine compounds, comprising nitroso (N (R1) (R2) -n=o) structures, are genotoxic substances for certain animals, some of which have been listed by the international agency for research on cancer (IARC) as possible or potential human carcinogens. These nitrosamine compounds, known as "group of interest" compounds in the human drug registration technical requirements international consortium (International Conference on Harmonization, abbreviated ICH) guide document M7 (R1), evaluate and control DNA reactive (mutagenic) impurities in pharmaceuticals to limit potential carcinogenic risk (2018, 3). The ICH guidelines suggest that any known mutagenic/carcinogen, including nitrosamine compounds, be controlled at intake levels where the risk of human cancer is negligible.

Currently, the U.S. Food and Drug Administration (FDA) has identified seven nitrosamine impurities that may theoretically exist in pharmaceutical products because of the use of manufacturing processes and materials that may lead to the formation of nitrosamines: n, N-dimethylnitrosamine (NDMA), N-Nitrosodiethylamine (NDEA), N-nitroso-N-methyl-4-aminobutyric acid (NMBA), N-Nitrosoisopropylethylamine (NIPEA), N-Nitrodiisopropylamine (NDIPA), N-Nitrodibutylamine (NDBA) and N-Nitrosomethylaniline (NMPA). Among them, five impurities (NDMA, NDEA, NMBA, NIPEA and NMPA) have been actually detected in crude drugs or medicines.

For example, some preliminary results of FDA experiments indicate that NDMA levels in certain ranitidine products have exceeded acceptable levels. In recent years, the FDA has also found that metformin products contain NDMA impurities, and that NDMA levels are detected in certain batches above the FDA recommended acceptable intake limit.

At present, some studies have attempted to control and reduce the formation of these nitrosamine impurities by technical means, such as: CN109748905a uses hydrogen peroxide to replace sodium nitrite to reduce the content of NDMA and NDEA in sartan drug substance; CN111686087a adopts a powder mixing direct compression process and a gastric-soluble film coating premix for coating, so that excessive heat is avoided in the production process, and the generation risk of NDMA is reduced; CN113081990a controls the genotoxic impurity N-Nitrosodimethylamine (NDMA) by controlling the content of impurity dimethylamine in the raw material drug metformin hydrochloride and the content of impurity nitrite in the auxiliary material hypromellose.

In particular with respect to the present invention,

there may also be problems with exceeding the nitrosamine impurity levels for valicarb or pharmaceutically acceptable salts, crystalline forms, solvates, compositions, formulations and other pharmaceutical compounds thereof.

Thus, there is also a need to detect and control nitrosamine impurities in valicarb, or pharmaceutically acceptable salts, crystalline forms, solvates, compositions, formulations, and other pharmaceutical compounds thereof. In view of this, the present invention has been made.

Disclosure of Invention

Aiming at the problems of drug genotoxicity and/or safety existing in the prior art: how to reduce the genotoxicity of the valicarb isopropyl drug and improve the safety of the product; the invention provides a solution through continuous research and development: a pharmaceutical composition of a secondary amine compound (valicarb, or a pharmaceutically acceptable salt thereof) having a low nitrosamine impurity content is provided. According to the invention, the content of nitrosamine impurities in the bulk drug is controlled, and pharmaceutically acceptable acid is added into the pharmaceutical composition of the secondary amine compound, so that the generation of nitrosamine impurities in the preparation process and/or the storage process is effectively inhibited and reduced, the stability of the pharmaceutical composition of the secondary amine compound is improved, and the content of nitrosamine impurities due to toxicity is controlled at a lower level, so that the pharmaceutical composition of the secondary amine compound (valicarb or pharmaceutically acceptable salt thereof) meeting the safety requirements (for example, the limit requirements of FDA) is finally obtained.

The invention provides a pharmaceutical composition, which comprises the following components: secondary amine compounds and pharmaceutically acceptable acids; wherein the secondary amine compound is valicarb or a pharmaceutically acceptable salt thereof;

the nitrosamine impurity content of the pharmaceutical composition is not more than 7.5ppm; the nitrosamine impurity is

The active ingredient of the pharmaceutical composition is valicarb, and the pharmaceutical composition also comprises pharmaceutically acceptable auxiliary materials;

the pharmaceutically acceptable auxiliary materials comprise one or more than two of the following auxiliary materials (1) - (5);

(1) and (3) a diluent: the diluent is one or more selected from microcrystalline cellulose, silicified microcrystalline cellulose, calcium hydrophosphate (including anhydrous substance and hydrate), mannitol, copovidone, hydroxypropyl cellulose, hydroxypropyl methylcellulose, ethyl cellulose, starch, maltodextrin, agar and guar gum; preferably, the diluent comprises microcrystalline cellulose and/or anhydrous dibasic calcium phosphate; more preferably, the diluent is microcrystalline cellulose and anhydrous calcium hydrogen phosphate; still further preferably, the weight ratio of the effective component to the diluent is 1 (100 to 300); preferably 1 (150 to 250); more preferably 1 (184-195);

(2) And (3) a disintegrating agent: the disintegrating agent is one or more than two of croscarmellose sodium, sodium starch glycolate, crospovidone, partially pregelatinized starch, pregelatinized hydroxypropyl starch, sodium carboxymethylcellulose and calcium carboxymethylcellulose; preferably, the disintegrant is partially pregelatinized starch; more preferably, the weight ratio of the effective component to the disintegrant is 1 (1-10); preferably 1, (2-6); more preferably 1 (3.5-4.5); for example 1:4;

(3) glidant: the glidant is one or more than two selected from colloidal silicon dioxide, gas phase silicon dioxide, colloidal silicon dioxide, corn starch, talcum powder, calcium silicate, magnesium silicate, tricalcium phosphate and silicon hydrogel; preferably, the glidant is colloidal silicon dioxide; more preferably, the weight ratio of the effective component to the glidant is 1 (0.1-8); preferably 1 (0.2-5); more preferably 1 (0.5-2.5); most preferably 1 (0.8-2), such as 1:0.8, 1:1 or 1:2;

(4) and (3) a lubricant: the lubricant is one or more than two of stearic acid, stearate, talcum powder, mineral oil, malt, glyceryl monostearate, glyceryl benzoate, glyceryl palmitostearate, hydrogenated vegetable oil, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate and sodium lauryl sulfate; preferably, the lubricant is stearic acid or stearate, and the stearate is magnesium stearate or sodium stearate; more preferably, the weight ratio of the effective component to the lubricant is 1 (0.1-8); preferably 1 (0.2-5); more preferably 1 (0.5-2.5); most preferably 1 (1-2), such as 1:1, 1:1.4 or 1:2;

(5) A coating agent; preferably, the coating agent is an opadry coating agent, for example: a white, blue, yellow, red, green or orange opadry coating; further preferred isWhite and/or +.>Blue。

Further, the method comprises the steps of,

in any of the above aspects (pharmaceutical compositions), the secondary amine compound is a pharmaceutically acceptable salt of valicarb; preferably, the secondary amine compound is valicarb hydrochloride or valicarb tartrate.

Further, the method comprises the steps of,

in any of the above embodiments (pharmaceutical compositions), the molar ratio of the secondary amine compound to the pharmaceutically acceptable acid is 1 (0.01-50) (e.g., a molar ratio of 1:0.05, 1:0.1, 1:0.5, 1:1, 1:1.5, 1:2, 1:2.5, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, or 1:40, etc.); preferably, the molar ratio of the secondary amine compound to the pharmaceutically acceptable acid is 1 (1-20); more preferably, the molar ratio of the secondary amine compound to the pharmaceutically acceptable acid is 1 (2-10).

Further, the method comprises the steps of,

in any one of the above-mentioned aspects (pharmaceutical compositions), preferably, the active ingredient of the pharmaceutical composition is valicarb.

Further, the method comprises the steps of,

the pharmaceutically acceptable auxiliary materials comprise (1) to (4) or (1) to (5) at the same time.

Further, the method comprises the steps of,

in any one of the above-mentioned aspects (pharmaceutical compositions), the pharmaceutical composition is a solid preparation or a liquid preparation;

preferably, the solid formulation is a tablet, for example a plain tablet or a coated tablet.

Further, the method comprises the steps of,

in any of the above aspects (pharmaceutical compositions), the pharmaceutical composition is an oral formulation.

Further, the method comprises the steps of,

in any of the above aspects (pharmaceutical compositions), the pharmaceutical composition has a pH of 4 or less under the following test conditions: preferably, the pH is 1 to 4, for example 1, 1.5, 2, 2.5, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9 or 4, etc.;

when the pharmaceutical composition is a solid formulation, the test conditions are: mixing the pharmaceutical composition with water having a ph=6±0.1 to form a dispersion having a concentration of 20% (i.e., 0.2 g/ml), and testing the pH value obtained;

when the pharmaceutical composition is a liquid formulation, the test conditions are: the solvent of the liquid preparation is water with pH=6+/-0.1, the concentration of the liquid preparation is 20% (g/ml), and the pH value is obtained by testing;

The above pH test conditions are not limited to the pharmaceutical composition, and the pH obtained by the above test conditions is the same as the above or within the error range, and falls within the scope of the present invention.

Further, the method comprises the steps of,

in any one of the above aspects (pharmaceutical compositions), the pharmaceutically acceptable acid is a solid acid or a liquid acid;

preferably, the solid acid is selected from one or more than two of tartaric acid, succinic acid, maleic acid, fumaric acid, citric acid, malic acid, ascorbic acid, benzenesulfonic acid, oxalic acid, triphenylacetic acid, 1-hydroxy-2-naphthoic acid and 3-hydroxy-2-naphthoic acid;

the liquid acid is one or more than two of hydrochloric acid, sulfuric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, acetic acid, trifluoroacetic acid, propionic acid, methanesulfonic acid and trifluoromethanesulfonic acid;

more preferably, the pharmaceutically acceptable acid is a solid acid; further preferably, the pharmaceutically acceptable acid is tartaric acid (L- (+) -tartaric acid or D- (-) -tartaric acid).

Further, the method comprises the steps of,

in any one of the above technical schemes (pharmaceutical compositions), the pharmaceutical composition comprises the following components in parts by weight:

1.71 parts of secondary amine compound;

100-250 parts of diluent;

2-10 parts of disintegrating agent;

0-5 parts of glidant;

0-5 parts of lubricant;

0.01 to 15 portions of pharmaceutically acceptable acid.

Further, the method comprises the steps of,

in any of the above technical schemes (pharmaceutical compositions), the pharmaceutical composition comprises the following components in parts by weight:

1.71 parts of secondary amine compound;

100-250 parts of diluent;

2-10 parts of disintegrating agent;

0-5 parts of glidant;

0-5 parts of lubricant;

0.01 to 15 portions of pharmaceutically acceptable acid.

Further, the method comprises the steps of,

1.71 parts of secondary amine compound;

184.47 parts of a diluent;

4 parts of disintegrating agent;

0.6 part of glidant;

5 parts of a lubricant;

1.71 to 3.42 portions of pharmaceutically acceptable acid.

Further, the method comprises the steps of,

the pharmaceutical composition also comprises a coating agent;

preferably, the coating agent is an opadry coating agent (including but not limited to, white, blue, yellow, red, green or orange opadry coating agents); further preferred are White and/or Blue.

Further, the method comprises the steps of,

in any of the above technical schemes (pharmaceutical compositions), the pharmaceutical composition is any one of the following formulas:

Scheme one:

the pharmaceutical composition comprises the following components in parts by weight:

1.71 parts of valicarb isopropyl tartrate;

64 parts of anhydrous calcium hydrophosphate;

120.47 parts of microcrystalline cellulose;

4 parts of partially pregelatinized corn starch;

0.6 parts of colloidal silica;

5 parts of stearic acid;

3.42 parts of L- (+) -tartaric acid.

Scheme II:

1.71 parts of valicarb isopropyl tartrate;

64 parts of anhydrous calcium hydrophosphate;

120.47 parts of microcrystalline cellulose;

4 parts of partially pregelatinized corn starch;

0.6 parts of colloidal silica;

5 parts of stearic acid;

1.71 parts of L- (+) -tartaric acid;

scheme III:

1.71 parts of valicarb isopropyl tartrate;

64 parts of anhydrous calcium hydrophosphate;

120.47 parts of microcrystalline cellulose;

4 parts of partially pregelatinized corn starch;

0.6 parts of colloidal silica;

5 parts of stearic acid;

3.42 parts of L- (+) -tartaric acid;

6 parts of Opadry-white (Opadry white premix) or Opadry-blue (Opadry blue premix).

Scheme IV:

1.71 parts of valicarb isopropyl tartrate;

64 parts of anhydrous calcium hydrophosphate;

120.47 parts of microcrystalline cellulose;

4 parts of partially pregelatinized corn starch;

0.6 parts of colloidal silica;

5 parts of stearic acid;

1.71 parts of L- (+) -tartaric acid;

Further, the method comprises the steps of,

in any of the above-described aspects (pharmaceutical compositions), the pharmaceutical composition has a nitrosamine impurity content of no greater than 7.4ppm, no greater than 7.3ppm, no greater than 7.2ppm, no greater than 7.1ppm, no greater than 7.0ppm; no more than 6.9ppm, no more than 6.8ppm, no more than 6.7ppm, no more than 6.6ppm, no more than 6.5ppm, no more than 6.4ppm, no more than 6.3ppm, no more than 6.2ppm, no more than 6.1ppm, no more than 6.0ppm; no more than 5.9ppm, no more than 5.8ppm, no more than 5.7ppm, no more than 5.6ppm, no more than 5.5ppm, no more than 5.4ppm, no more than 5.3ppm, no more than 5.2ppm, no more than 5.1ppm, no more than 5.0ppm; no more than 4.9ppm, no more than 4.8ppm, no more than 4.7ppm, no more than 4.6ppm, no more than 4.5ppm, no more than 4.4ppm, no more than 4.3ppm, no more than 4.2ppm, no more than 4.1ppm, no more than 4.0ppm, no more than 3.9ppm, no more than 3.8ppm, no more than 3.7ppm, no more than 3.6ppm, no more than 3.5ppm, no more than 3.4ppm, no more than 3.3ppm, no more than 3.2ppm, no more than 3.1ppm, no more than 3.0ppm; no more than 2.9ppm, no more than 2.8ppm, no more than 2.7ppm, no more than 2.6ppm, no more than 2.5ppm, no more than 2.4ppm, no more than 2.3ppm, no more than 2.2ppm, no more than 2.1ppm, or no more than 2.0ppm.

The invention also provides a preparation method of the pharmaceutical composition, which comprises the following steps:

mixing all the components, granulating, tabletting, and optionally coating;

preferably, the method comprises the steps of,

the preparation method comprises the following steps: colloidal silicon dioxide, anhydrous calcium hydrophosphate, valicarb tartrate and the pharmaceutically acceptable acid are sieved by a 40-mesh sieve, uniformly mixed, added with microcrystalline cellulose, part of pregelatinized corn starch and part of stearic acid in grains, uniformly mixed, granulated, added with part of stearic acid outside grains, uniformly mixed, tabletted and optionally coated to obtain the compound calcium phosphate;

more preferably, the process is carried out,

the granulating is dry granulating or wet granulating;

the tabletting is a direct tabletting method.

The invention also provides application of the pharmaceutical composition in the preparation of medicines; the medicament is used for treating and/or preventing the following diseases: inflammatory bowel disease, ulcerative colitis, pyoderma gangrenosum, crohn's disease, irritable bowel syndrome, spasmodic dystonia, chronic pain, acute pain, diarrhea, cystitis, vasoconstriction, anxiety, mania, depression, bipolar disorder, autism, sleep disorder, jet lag, amyotrophic lateral sclerosis, cognitive impairment Known dysfunction, cognitive impairment caused by alcohol, barbiturates, vitamin deficiency, recreational drugs, lead, arsenic or mercury, alzheimer's disease, senile dementia, vascular dementia, parkinson's disease, multiple sclerosis, AIDS, encephalitis, trauma, hepatorenal encephalopathy, hypothyroidism, pick's disease, coltsaokoff's syndrome, forehead dementia or subcortical dementia, hypertension, bulimia, anorexia, obesity, cardiac arrhythmia, gastric acid hypersecretion, ulcers, pheochromocytomas, progressive supranuclear palsy, nicotine, tobacco products, alcohol, benzodiazepinesChemical dependence and addiction to barbiturates, opioids or cocaine, headache, migraine, stroke, traumatic brain injury, obsessive-compulsive disorder, psychosis, huntington's chorea, tardive dyskinesia, hyperkinesia, dyskinesia, schizophrenia, multi-infarct dementia, age-related cognitive decline, epilepsy (seizure deficiency), attention deficit hyperactivity disorder or tourette's syndrome;

preferably, the medicament meets one of the following conditions a to c:

a. the medicine is used for treating nicotine dependence, addiction or withdrawal;

b. The medicine is used for treating dependence, addiction or withdrawal of tobacco products;

c. the medicine is a smoking cessation medicine.

Another object of the invention is to provide a new application (use).

The specific technical proposal is that,

use of a pharmaceutically acceptable acid to inhibit or slow down the nitrosation of a secondary amine compound (nitrosation refers to the nitrosation of a secondary amine structure to produce a nitrosamine structure); wherein the secondary amine compound is valicarb or a pharmaceutically acceptable salt thereof;

preferably, the use of the pharmaceutically acceptable acid in inhibiting or slowing down the nitrosation of a secondary amine compound satisfies one or more of the following conditions Ia to Id (e.g., satisfies condition Ia alone, condition Ib alone, condition ic alone, etc.):

the molar ratio of la, the secondary amine compound, and the pharmaceutically acceptable acid is from 1:0.01 to 50 (e.g., a molar ratio of 1:0.05, 1:0.1, 1:0.5, 1:1, 1:1.5, 1:2, 1:2.5, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, or 1:40, etc.);

the secondary amine compound Ib is a pharmaceutically acceptable salt of valicarb;

ic, the pharmaceutically acceptable acid is a solid acid or a liquid acid; preferably, the solid acid is selected from one or more than two of tartaric acid, succinic acid, maleic acid, fumaric acid, citric acid, malic acid, ascorbic acid, benzenesulfonic acid, oxalic acid, triphenylacetic acid, 1-hydroxy-2-naphthoic acid and 3-hydroxy-2-naphthoic acid; the liquid acid is one or more than two of hydrochloric acid, sulfuric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, acetic acid, trifluoroacetic acid, propionic acid, methanesulfonic acid and trifluoromethanesulfonic acid;

Id. The pharmaceutically acceptable acid is added in an amount such that the pH value of the mixture X is less than or equal to 4 under the following test conditions; the mixture X comprises the pharmaceutically acceptable acid and the secondary amine compound, preferably at a pH of 1 to 4, for example 1, 1.5, 2, 2.5, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9 or 4, etc.; when the mixture is solid, the test conditions are: mixing the mixture with water having a ph=6±0.1 to form a dispersion having a concentration of 20% (g/ml), and testing the pH value obtained; when the mixture is liquid, the test conditions are: the solvent of the liquid is water with pH=6+/-0.1, the concentration of the liquid is 20% (g/ml), and the pH value is obtained by testing;

the above pH test conditions are not limited to the mixture, and the pH obtained by the above test conditions is the same as the above or within the error range, and falls within the scope of the present invention.

More preferably, the use of the pharmaceutically acceptable acid in inhibiting or slowing down the nitrosation of a secondary amine compound satisfies one or more of the following conditions IIa to IIc (e.g., satisfies condition IIa alone, condition IIb alone, condition IIc alone, etc.):

The molar ratio of IIa, secondary amine compound and pharmaceutically acceptable acid is 1 (1-20); further preferably, the molar ratio of secondary amine compound to pharmaceutically acceptable acid is 1 (2-10);

IIb, the secondary amine compound is valicarb hydrochloride or valicarb tartrate;

IIc, wherein the pharmaceutically acceptable acid is a solid acid; further preferably, the pharmaceutically acceptable acid is tartaric acid (L- (+) -tartaric acid or D- (-) -tartaric acid);

it is further preferred that the composition comprises,

the pharmaceutically acceptable acid is used for inhibiting or slowing down the nitrosamine of the secondary amine compound and simultaneously satisfies the conditions Ia-Id or IIa-IIc.

Further, the use of the pharmaceutically acceptable acid to inhibit or slow down the nitroamination of a secondary amine compound refers to inhibiting or slowing down the nitroamination of the secondary amine compound

The invention also provides a method for inhibiting or slowing down the nitrosamine of the secondary amine compound, which specifically comprises the following steps: uniformly mixing a secondary amine compound or a composition thereof with pharmaceutically acceptable acid; wherein the secondary amine compound is valicarb or a pharmaceutically acceptable salt thereof.

Further, the method for inhibiting or slowing down the nitrosamine of the secondary amine compound comprises the following steps: adding pharmaceutically acceptable acid into secondary amine compound or its composition, and mixing.

Further, the composition of secondary amine compounds comprises all components of the pharmaceutical composition of any of the preceding claims except pharmaceutically acceptable acids.

Further, the method for inhibiting or slowing down the nitrosamine of the secondary amine compound satisfies one or more of the following conditions Ia to ic (for example: satisfies condition Ia alone, condition Ib alone, condition ic alone, etc.):

the molar ratio of la, secondary amine compound, and pharmaceutically acceptable acid is 1:0.01-50 (e.g., molar ratio of 1:0.05, 1:0.1, 1:0.5, 1:1, 1:1.5, 1:2, 1:2.5, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, or 1:40, etc.);

ic, the pharmaceutically acceptable acid is a solid acid or a liquid acid; the solid acid is selected from one or more than two of tartaric acid, succinic acid, maleic acid, fumaric acid, citric acid, malic acid, ascorbic acid, benzenesulfonic acid, oxalic acid, triphenylacetic acid, 1-hydroxy-2-naphthoic acid and 3-hydroxy-2-naphthoic acid; the liquid acid is one or more than two of hydrochloric acid, sulfuric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, acetic acid, trifluoroacetic acid, propionic acid, methanesulfonic acid and trifluoromethanesulfonic acid;

Id. The pharmaceutically acceptable acid is added in an amount such that the pH value of the mixture X is less than or equal to 4 under the following test conditions; the mixture X comprises the pharmaceutically acceptable acid and the secondary amine compound, preferably at a pH of 1 to 4, for example 1, 1.5, 2, 2.5, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9 or 4, etc.; when the mixture is solid, the test conditions are: mixing the mixture with water with the pH value of 6+/-0.1 to form a dispersion liquid with the concentration of 20%, and testing the obtained pH value; when the mixture is liquid, the test conditions are: the solvent of the liquid is water with pH=6+/-0.1, the concentration of the liquid is 20%, and the pH value obtained by testing is measured;

preferably, the method of inhibiting or slowing down the nitrosamine of a secondary amine compound satisfies one or more of the following conditions IIa to IIc (e.g., satisfies condition IIa alone, condition IIb alone, condition IIc alone, etc.):

it is further preferred that the composition comprises,

the method for inhibiting or slowing down the nitrosamination of the secondary amine compound simultaneously meets the conditions Ia-Id or IIa-IIc.

Further, the method for inhibiting or slowing down the nitroamination of the secondary amine compound means that the nitroamination of the secondary amine compound is inhibited or slowed down

On the basis of conforming to the common knowledge in the field, the above preferred conditions can be arbitrarily combined to obtain the preferred examples of the invention.

The reagents and materials used in the present invention are commercially available.

The beneficial effects of the invention are mainly as follows: according to the invention, the nitrosamine impurity content in the bulk drug is controlled (namely, the valicarb acid bulk drug with low nitrosamine impurity content is adopted), and the pharmaceutically acceptable acid is added into the pharmaceutical composition of the secondary amine compound (valicarb acid or pharmaceutically acceptable salt thereof), so that the generation of nitrosamine impurities in the preparation process and/or the storage process is effectively inhibited and reduced, the stability of the pharmaceutical composition of the secondary amine compound is improved, the content of the nitrosamine impurities due to toxicity is controlled at a lower level, and the pharmaceutical composition of the secondary amine compound (valicarb acid or pharmaceutically acceptable salt thereof) with low nitrosamine impurity content is obtained, thereby meeting the safety requirements and the regulatory requirements of pharmaceutical products (for example, the limit requirements of FDA) and better guaranteeing the medication safety of patients.

It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.

Detailed Description

The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.

Chinese patent application CN2021109612747 is incorporated herein by reference in its entirety.

Example 1

Table 1, formulation A (1 mg) and its content

Wherein,

the 1.71 mg/tablet of valicarb-isopropyl tartrate can be converted into 1 mg/tablet of valicarb-isopropyl, namely: each tablet contains 1mg of active ingredient (valicarb), which can also be expressed as: 1mg of plain tablet (or 1mg of coated tablet, etc.). Whereas a 0.5mg tablet is one in which the amounts of the components are halved on the basis of a 1mg tablet. In the above formulation microcrystalline cellulose and anhydrous dibasic calcium phosphate are used as diluents, partially pregelatinized corn starch is used as a disintegrant, colloidal silicon dioxide is used as a glidant, stearic acid is used as a lubricant, L- (+) -tartaric acid is used as an acidity regulator (alternatively referred to as a pH regulator), and purified water is used as a coating solvent.

The preparation method (13 ten thousand tablets per batch) comprises the following steps:

multi-stage mixing

a. Sieving colloidal silica (Part I) and anhydrous calcium hydrogen phosphate (Part I) with a 40-mesh sieve, sieving with a discharger (diameter 1.2mm, speed 400+ -10 r/min), and stirring in a stirrer (rotation speed: 15 r/min) for 2min;

b. sieving valicarb tartrate with 40 mesh sieve, sieving with discharger (diameter 1.2mm, speed 800+ -15 r/min), and adding into the above mixer for continuous stirring for 10min;

c. the colloidal silicon dioxide (Part II), anhydrous calcium hydrophosphate (Part II) and L- (+) -tartaric acid are sieved by a 40-mesh sieve, sieved by a discharger (with the diameter of 1.2mm and the speed of 400+/-15 r/min), and then added into the stirrer for continuous stirring for 10min;

d. microcrystalline cellulose, partially pregelatinized corn starch, stearic acid (intra-granular fraction) were sieved through a discharger (diameter 1.2mm, speed 100.+ -. 5 r/min), and then added to the above-mentioned stirrer for continuous stirring for 10min.

Dry granulation

The mixture is granulated by using a dry granulator LGS120, and specific parameters are as follows: and (3) screening: 1.0mm; feed rate: 20-30 rpm; roller speed: 10+ -2 rpm; roll gap: 0.5-3.0 mm; oil pressure: 60-70 bar; rotational speed: 100-140 rpm.

Third and final mixing

Adding the granulated granules and stearic acid (extra granular part) into a stirrer (rotating speed: 15 r/min) and stirring for 10min to obtain a final mixture.

Fourth, tabletting (1 mg plain tablet A)

The final blend was tabletted and the specific parameters are shown in table 2.

TABLE 2 main process parameters of tabletting

Fifthly, coating (1 mg white coating tablet A)

Adding the Opadry-white (Opadry white premix) into purified water according to the proportion, and uniformly mixing to obtain a coating film dispersion liquid. The tablet (plain tablet) is coated by a coating machine, and the specific parameters are as follows: pan speed (Pan speed): 3-12 r/min; inlet/outlet air speed (Inlet/outlet air rotating speed): 1200/1600rpm; intake air temperature (Inlet airtemperature): 60 ℃; start spray gun pressure (Starting spray gun pressure): 2.5.+ -. 0.5bar; atomization pressure (atom izationpresure): 2.0.+ -. 0.5bar; latex tube diameter (Latex tube diameter): 4.00-5.00 mm; peristaltic pump speed (Peristaltic pump rotating speed): 20-50 r/min.

The coating agent was replaced with opadry-blue (opadry blue premix) and 1mg of blue coated tablet a was prepared under the same conditions.

Sixth, packaging by aluminum plastic.

Example 2

Formula B (specification 1 mg): the dosage of L- (+) -tartaric acid is changed to 1.71 mg/tablet, and other conditions are unchanged. According to the same production method as in example 1, 1mg of plain tablet B, 1mg of white coated tablet B and 1mg of blue coated tablet B were produced.

Example 3

Formula Y (specification 1 mg): l- (+) -tartaric acid is not added into the tablet, and other conditions are not changed. According to the same production method as in example 1, 1mg of plain tablets Y, 1mg of white coated tablets Y and 1mg of blue coated tablets Y were produced.

Example 4

The prepared valicarb tartrate tablet was dispersed in purified water (ph=6.01 of purified water, as measured), to form a dispersion having a concentration of 20% (g/ml) (indicating that 100ml of the solution or dispersion contains 20g of valicarb tartrate tablet), and after 5 minutes of sonication, the pH was measured. The nitrosamine impurity P08 content in the valicarb tablet was detected by HPLC Q-exact-MS/MS (Vanquish HPLC Q-exact-MS/MS).

The results are shown in tables 3 to 5.

Table 3, pH and detection results of nitrosamine impurity P08 content (Tablet Core Tablet)

Table 4, pH and detection results of nitrosamine impurity P08 content (White Coating tablet)

Table 5, pH and detection results of nitrosamine impurity P08 content (Blue Coating tablet)

Wherein,

API (Active Pharmaceutical Ingredient): refers to a valicarb tartrate bulk drug.

Initially: refers to the detection immediately after the valicarb tartrate tablet is prepared.

High temperature test: after the valicarb tartrate tablet is prepared, the valicarb tartrate tablet is placed for a period of time at 60 ℃ and then is detected (refer to: chinese pharmacopoeia).

Symbol ×: represents a liquid sample after the pH adjustment, and the liquid sample is detected.

Symbol #: indicating a liquid sample after the pH adjustment, performing a high temperature test, and then detecting the liquid sample.

Example 5

The HPLC Q-exact-MS/MS (Vanquish HPLC Q-exact-MS/MS) method was used for qualitative and/or quantitative detection of nitrosamine impurity P08 in valicarb tablets, and was performed and validated by methodology, the detection results are specified in example 4.

The HPLC Q-actual-MS/MS chromatographic conditions and mass spectrometry conditions are shown in Table 6.

TABLE 6 chromatographic conditions and Mass Spectrometry conditions for HPLC Q-actual-MS/MS

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1. Solution preparation

Control stock solution: 7.5mg of nitrosamine compound P08 (purity not less than 98%) is weighed into a 100ml measuring flask, and acetonitrile is added for dissolution, and the volume is fixed (fixed solution 1A). 1.0ml of the constant solution 1A was placed in a 100ml measuring flask, and acetonitrile was added to the flask to fix the volume (constant solution 1B). 1.0ml of the fixed solution 1B is taken and placed in a 20ml measuring flask, acetonitrile is added for constant volume, and a reference stock solution is obtained.

Control solution: taking 1.0ml of reference substance stock solution, placing the stock solution into a 10ml measuring flask, and adding acetonitrile to fix the volume to obtain the reference substance solution.

Sample solution: 10 pieces of valicarb acid tablet are taken and placed in a 10ml measuring flask, 3ml of purified water and 3ml of acetonitrile are added, ultrasound is carried out for 30s, the mixture is diluted to scale by acetonitrile, and the mixture is uniformly mixed.

Sample addition of the labeling solution: 10 pieces of valicarb acid vandula angustifolia tablets are taken and placed in a 10ml measuring flask, 3ml of purified water and 3ml of acetonitrile are added, ultrasound is carried out for 30s, then 1.0ml of reference stock solution is added, acetonitrile is added to fix the volume to scale, and the mixture is uniformly mixed.

Placebo solution: 1g of placebo (corresponding to the tablet with the active ingredient removed) was weighed into a 10ml measuring flask, 3ml of purified water and 3ml of acetonitrile were added, sonicated for 30s, diluted to scale with acetonitrile, and mixed well.

Placebo labeling solution: 1g of placebo is weighed and placed in a 10ml measuring flask, 3ml of purified water and 3ml of acetonitrile are added, ultrasound is conducted for 30s, 1.0ml of control stock solution is added, acetonitrile is added for constant volume dilution to scale, and the mixture is uniformly mixed.

Blank: acetonitrile.

2. Specialization of

Taking blank, reference substance solution, sample solution, placebo solution, sample labeling solution and placebo labeling solution, and respectively carrying out sample injection analysis.

The results show that the retention time of the target objects of the reference substance solution, the sample solution and the standard adding solution is about 7.80min, no obvious interference exists at the peak position of the target objects, the requirement of the separation degree is met, and the specificity is good.

3. Solution stability

Taking a control solution and a sample solution, placing at room temperature, and carrying out sample injection analysis at intervals: 0. the peak area ratio (i.e., peak area change rate) of each time point to zero point was calculated for 2, 4, 6, 8, 10, 12 hours.

The results show that the peak area change rate of the nitrosamine compound P08 in the control solution is less than 12% in 12 hours, and the peak area change rate of the nitrosamine compound P08 in the sample solution is less than 3.5% in 12 hours.

4. System applicability

And the control solution is taken and analyzed for 5 times, the RSD of the peak area of the target object is calculated, the result is 1.41%, and the system applicability is good.

5. Repeatability of

6 sample solutions were prepared in parallel and analyzed by sample injection. The result shows that the RSD of the impurity compound P08 content in 6 parts of sample solution is 3.49%, meets the acceptance standard, and has good repeatability.

6. Intermediate precision

Two analysts weighed 6 sample solutions at different times, sample analysis was performed separately, and the average and RSD were calculated. The results showed that rsd=5.93%.

7. Linearity of

Respectively preparing reference substance solutions of 0.015ng/ml, 0.38ng/ml, 1.88ng/ml, 3.77ng/ml, 5.65ng/ml, 18.83ng/ml, 37.66ng/ml and 753.2ng/ml, and carrying out sample injection analysis; taking the concentration (X) of the target object as an abscissa and the peak area (Y) as an ordinate, carrying out linear regression, and calculating to obtain a linear equation and a linear correlation coefficient r;

The results show that the linear equation: y=462785x+1000000, the linear correlation coefficient r=0.9999, and good linearity in the concentration range of 0.015ng/ml to 753.2 ng/ml.

8. Accuracy of

The result shows that the recovery rate of the sample adding standard solution is between 85% and 95%, the RSD is 2.03%, the sample adding standard meets the acceptance standard, and the accuracy is good.

9. Detection Limit (LOD)

Lod=13.2 ppb, the average S/N (signal to noise ratio) of the target peak is 75.94, the requirement of greater than 3 is met, and the method is sensitive.

10. Quantitative Limit (LOQ)

Loq=17.6 ppb, the average S/N of the target peak is 103.35, meeting the requirement of greater than 10, the method is sensitive.

11. Durability of

When the temperature change of the chromatographic column is +/-2 ℃, the result shows that the method has good durability.

The nitrosamine impurity compound P08 content in the sample was calculated according to the following formula:

c (ppm) =p 08 concentration in sample solution (ng/ml)/valicarb tartrate concentration in sample solution (mg/ml).

Example 6

Preparation of nitrosamine Compound P08

To a 100mL three-necked flask, 4.0g of Compound M02 (valicarb) and 45mL of Tetrahydrofuran (THF) were added, and the solution was stirred; 8.2mL of NaNO was added ₂ The aqueous solution (concentration 0.4 g/mL) was added with 2.9g of acetic acid, the temperature was raised to 52℃and the TLC was monitored for the completion of the reaction, and a solid was precipitated, filtered and dried under vacuum to give nitrosamine compound P08 having a purity of 98.87% (area normalization method).

Nitrosamine Compound P08 ¹ H-NMR、 ¹³ C-NMR and IR spectra are shown in tables 7 to 9, respectively; M+H ⁺ = 241.20; UV: the maximum absorption wavelength in acetonitrile solution was 204.40nm.

TABLE 7 nitrosamine Compound P08 ¹ H-NMR spectrum

TABLE 8 nitrosamine Compound P08 ¹³ C-NMR spectrum

Table 9 IR spectrum of nitrosamine Compound P08

Absorption wave number (cm) ^-1 )	Attribution to
		3048.8	Ar-H
2961.4,2931.8	Saturated C-H stretching vibration
		1925.2	Benzene ring c=c stretching vibration
1575.2	N=o stretching vibration
		885.2	Bending vibration of benzene ring C-H

Example 7

The ultra-high performance liquid chromatography triple quadrupole mass spectrometry (UPLC-MS/MS) is used for qualitative and/or quantitative detection of nitrosamine impurities (compound P08) in valicarb-cymbidium tartrate bulk drug, and is performed and passes methodological verification.

The chromatographic conditions and mass spectrometric conditions of UPLC-MS/MS are shown in Table 10.

TABLE 10 chromatographic conditions and Mass Spectrometry conditions for UPLC-MS/MS

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1. Solution preparation

A diluent: purified water

Control stock solution: 19.970mg of nitrosamine compound P08 (purity not less than 98%) is weighed, placed in a 10ml measuring flask, 8ml of acetonitrile is added, then diluent is added for dissolution and dilution to scale, and uniformly mixed.

Reference mother liquor: the control stock solution was measured at 37.5 μl in a 10ml measuring flask, diluted to scale with diluent, and mixed well.

2. System applicability

System applicability solution (100% limit level control solution): about 100.0 μl of the control mother liquor is measured and placed in a 10ml measuring flask, and the diluent is diluted to the scale and mixed well.

The RSD of the peak area of the target in the 6-needle system applicability solution was calculated by continuously feeding the 6-needle system applicability solution, and as a result, the system applicability was good.

3. Linearity of

Standard curve solution: respectively weighing 50.0 μl, 100.0 μl, 150.0 μl and 200.0 μl of reference mother liquor, placing into 4 10ml measuring flasks, diluting with diluent to scale, mixing, and sequentially taking as linear solutions L-3, L-4, L-5 and L-6. Weighing the L-3 solution 800 mu 1 to 10ml measuring flask, diluting the diluent to the scale, and uniformly mixing to obtain the L-2. Weighing the L-2 solution 1000 mu 1 to 10ml measuring flask, diluting the diluent to the scale, and uniformly mixing to obtain the L-1.

Taking standard curve solutions for sample injection analysis respectively, taking the concentration (X) of a target object as an abscissa (unit: ng/ml), taking the peak area (Y) as an ordinate, and carrying out linear regression, so as to obtain a linear equation and a linear correlation coefficient r; the results show that the linear equation: y=7370X-12900, the linear correlation coefficient r=0.999, the linearity is good, and the linearity is good in a concentration range of 0.2951ng/ml to 147.6ng/ml (namely: 0.02951ppm to 14.76 ppm).

4. Detection Limit (LOD)

LOD solution: weighing the L-1 solution into a 5000 mu 1-10 ml measuring flask, diluting the diluent to a scale, and uniformly mixing to obtain the LOD solution.

Taking LOD solution to continuously sample for 2 needles, wherein the S/N (signal to noise ratio) of a target peak is respectively 11 and 8, so that the requirement of not less than 3 is met, and the method is sensitive; lod= 0.01476ppm.

5. Quantitative Limit (LOQ)

Taking a quantitative limiting solution (L-1 solution) for continuous sampling for 6 needles, wherein the S/N of a target peak is 23, 24, 17, 12, 24 and 14 respectively, so that the requirement of not less than 10 is met, and the method is sensitive; loq= 0.02951ppm.

6. Accuracy of

Sample solution: about 100mg of the sample (e.g., valicarb, purchased or synthesized) is weighed into a 10ml measuring flask, diluted to a scale with a diluent, and mixed well. 2 parts were prepared in parallel.

Accuracy solution: about 100mg of the sample is weighed and placed in a 10ml measuring flask, 50.0 mu l (50% limit level), 100.0 mu l (100% limit level) and 150.0 mu l (150% limit level) of reference mother liquor are respectively added, and diluted to a scale by a diluent, and the mixture is uniformly mixed to obtain the finished product. 3 replicates were prepared for each level of accuracy.

The result shows that the recovery rate of the solution at the 50% limit level accuracy is between 95% and 102%, and the RSD is 4%; the recovery rate of the solution is between 90% and 103% at the 100% limit level accuracy, and the RSD is 7%; the recovery rate of the solution at the 150% limit level accuracy is 98% -100%, the RSD is 1%, the solution meets the acceptance standard, and the accuracy is good.

7. Repeatability of

About 100mg of the sample is weighed and placed in a 10ml measuring flask, 100.0 μl of reference mother liquor is added, the diluent is diluted to scale, and the mixture is uniformly mixed. 6 parts were prepared in parallel.

The results showed that the RSD of the nitrosamine compound P08 content in the 6 parts of the repetitive solution was 6%, which meets the acceptance criteria and the reproducibility was good.

8. Specialization of

Blank solution (diluent), 100% limit level control solution, sample solution (same as "repeated" sample solution), 100% limit level accuracy solution were taken and analyzed separately.

The results show that the retention time of the target object of the sample solution, the 100% limit level reference substance solution and the 100% limit level accuracy solution is 3.47min, no obvious interference exists at the peak position of the target object, the requirement of the separation degree is met, and the specificity is good.

9. Intermediate precision

Intermediate precision solution: about 100mg of the sample is weighed and placed in a 10ml measuring flask, 100.0 μl of reference mother liquor is added, the diluent is diluted to scale, and the mixture is uniformly mixed. 6 parts were prepared in parallel. And 6 parts of "7, repetitive" solutions were added.

The concentration RSD of the total 12 parts of the intermediate precision solution and the 6 parts of the repeated solution is calculated to be 9%, the requirement of not more than 15% is met, the acceptance standard is met, and the intermediate precision is good.

10. Solution stability

And placing the 100% limit horizontal accuracy solution at 15 ℃, analyzing at intervals, and calculating the concentration ratio of each time point to the zero point, wherein the ratio is 96-113% in 21 h.

11. Durability of

And 2 needles are respectively injected under each condition when the temperature change of the chromatographic column is +/-2 ℃ and the initial proportion of the organic phase is +/-1% and the flow rate is +/-0.01 ml/min, so that the concentration value of the target in the solution with the 100% limit horizontal accuracy is calculated, the RSD of the concentration value of the target in 6 parts of solution in the fluctuation of the same parameter is between 4% and 6%, and the method has good durability.

The content of nitrosamine compound P08 in the sample was calculated, detected 2 times, and averaged according to the following formula:

p08 content: c (ppm) =P 08 concentration in sample solution (ng/ml)/[ m (mg)/V (ml) ]

m: sample mass, mg;

v: the samples were diluted in volume, ml.

Example 8

Whether nitrosamine impurity compound P08 is genotoxic (capable of mutation or carcinogenesis) is detected.

1 materials and methods

1.1 Experimental strains

Salmonella typhimurium TA97a, TA98, TA100, TA102, TA1535, from: MOLT OX, usa, available from Shanghai, biotechnology, inc. The test was performed using an identified desirable Salmonella typhimurium strain.

1.2 Metabolic activation System

Rat liver S9, source: shanghai Bao Ji Biotech Co., ltd.

1.3 test article

Name: nitrosamine impurity compound P08, trait: white powder.

1.4 major instrumentation and reagents

X SR-204 electronic balance (Metrehler, switzerland); high pressure steam sterilizer (SANYO Co., japan); a water-proof constant temperature incubator (Shanghai-constant technology Co., ltd.); stuart temperature controlled shaker incubator (Stuart Co., UK); BHC-1300IIA2 biosafety cabinet (Suzhou Antai air technologies Co., ltd.).

Glucose 6-phosphate, source: beijing carboline technologies Co., ltd; coenzyme II, source: national pharmaceutical group chemical agents, inc; nutrient broth, source: beijing land bridge technology Co., ltd; technical agar powder, source: guangdong Crypton microorganism technologies Co., ltd; sterilized water for injection, the source: chenxin pharmaceutical industry stock limited; dixon, source: CHEMSERVICE; methyl methanesulfonate, source: beijing carboline technologies Co., ltd; 2-aminofluorene, source: shanghai Crystal pure reagent Co., ltd; 1, 8-dihydroxyanthraquinone, source: SIGMA-ALDRICH Co., USA; cyclophosphamide, source: SIGMA-ALDRICH Co., USA; sodium azide, source: RIEDEL-SEELIE.

1.5 test methods (Ames test plate incorporation method)

The experiment used a plate incorporation method.

Dose selection and test formulation of 5.l

Dose design: the experiments were performed at 5mg, 2.5mg, 1.25mg, 0.625mg, 0.3125mg, 0.15625mg per dish.

Test object preparation: the product is prepared into 50mg/ml solution by dimethyl sulfoxide, and is diluted into 25mg/ml, 12.5mg/ml, 6.25mg/ml, 3.125mg/ml and 1.5625mg/ml by dimethyl sulfoxide in sequence.

1.5.2 test methods.

The cryopreserved bacterial solutions TA97a, TA98, TA100, TA102 and TA15350.1ml were inoculated into 10ml of nutrient broth medium, respectively, and cultured at 37℃with shaking (100 times/min) for 10 hours. 2.0ml of the top medium containing 0.5mmol/L histidine-0.5 mmol/L biotin is packaged in test tubes, sterilized at 0.103MPa for 20min and incubated in a water bath at 50 ℃. Then each tube is added with 0.1ml of the test solution and S in turn ₉ Mixing 0.5ml of the mixed solution (when metabolic activation is needed) and 0.1ml of the bacterial solution, fully and uniformly mixing, rapidly pouring the mixed solution into a bottom agar plate, and rotating the plate to uniformly distribute the mixed solution. After being horizontally placed and solidified by condensation, the mixture is inverted and incubated in an incubator at 37 ℃ for 48 hours, and the number of the colony after transformation of each dish is counted. Three parallel plates were made for each dose, and self-priming, solvent control, and positive control were set simultaneously.

1.6 statistics of test data

Data are expressed as mean ± standard deviation (x ± s), and data analysis calculations are performed using SPSS 19.0.

1.7 result determination

If the number of the revertant colonies of the test object is twice or more than that of the solvent control revertant colonies and is in a dose-response relationship, the test object is judged to be mutation positive; the test agent is judged to be mutationally positive if the test agent has a positive response and is repeatable at any dose. If the test substance is determined by the above test strain, then either S is added ₉ And not adding S ₉ All negative under the conditions, the test object is mutation negative.

2 test results

The results are shown in Table 11.

Table 11, ames test results (x.+ -. S)

Continuous table Ames test results (x+ -s)

Ames test results show that in the absence of S ₉ Under the conditions of (2) the colony count of TA1535 was more than twice that of the solvent control group at 625. Mu.g/dish of nitrosamine impurity compound P08. At the addition of S ₉ At doses of 2500 μg/dish, the number of TA98 colonies exceeded twice that of the solvent control group, and at doses of 5000, 2500, 1250, 625 μg/dish, the number of TA1535 colonies exceeded twice that of the solvent control group, in a dose-response relationship. This indicates that nitrosamine impurity compound P08 is mutationally positive for Salmonella typhimurium with and without the addition of a metabolic activation system.

The invention is, of course, capable of other numerous embodiments and of being practiced in accordance with the invention and carried out by those skilled in the art without departing from the spirit and spirit of the invention, and it is intended that all such modifications and/or variations be regarded as being within the scope of the appended claims.

Claims

1. A pharmaceutical composition, characterized in that it comprises the following components: secondary amine compounds and pharmaceutically acceptable acids; wherein the secondary amine compound is valicarb or a pharmaceutically acceptable salt thereof;

2. The pharmaceutical composition of claim 1, wherein the secondary amine compound is a pharmaceutically acceptable salt of valicarb;

preferably, the secondary amine compound is valicarb hydrochloride or valicarb tartrate.

3. The pharmaceutical composition according to claim 1 or 2, wherein the molar ratio of the secondary amine compound and the pharmaceutically acceptable acid is 1 (0.01-50), e.g. the molar ratio is 1:0.05, 1:0.1, 1:0.5, 1:1, 1:1.5, 1:2, 1:2.5, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30 or 1:40;

preferably, the molar ratio of the secondary amine compound to the pharmaceutically acceptable acid is 1 (1-20); more preferably, the molar ratio of secondary amine compound to pharmaceutically acceptable acid is 1 (2-10).

4. The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutically acceptable excipients comprise (1) to (4) or (1) to (5) simultaneously.

5. The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is a solid or liquid formulation and/or the pharmaceutical composition is an oral formulation; preferably, the solid formulation is a tablet, for example a plain tablet or a coated tablet.

6. The pharmaceutical composition of claim 5, wherein the pharmaceutical composition has a pH of 4 or less under the following test conditions: preferably, the pH is 1 to 4, for example 1, 1.5, 2, 2.5, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9 or 4, etc.;

when the pharmaceutical composition is a solid formulation, the test conditions are: mixing the pharmaceutical composition with water having a ph=6±0.1 to form a dispersion having a concentration of 20%, and testing the pH value obtained;

when the pharmaceutical composition is a liquid formulation, the test conditions are: the solvent of the liquid preparation is water with pH=6+/-0.1, the concentration of the liquid preparation is 20%, and the obtained pH value is tested.

7. The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutically acceptable acid is a solid acid or a liquid acid;

preferably, the solid acid is selected from one or more than two of tartaric acid, succinic acid, maleic acid, fumaric acid, citric acid, malic acid, ascorbic acid, benzenesulfonic acid, oxalic acid, triphenylacetic acid, 1-hydroxy-2-naphthoic acid and 3-hydroxy-2-naphthoic acid; the liquid acid is one or more than two of hydrochloric acid, sulfuric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, acetic acid, trifluoroacetic acid, propionic acid, methanesulfonic acid and trifluoromethanesulfonic acid;

More preferably, the pharmaceutically acceptable acid is a solid acid; further preferably, the pharmaceutically acceptable acid is tartaric acid, e.g., L- (+) -tartaric acid or D- (-) -tartaric acid.

8. The pharmaceutical composition according to claim 4, wherein the pharmaceutical composition comprises the following components in parts by weight:

1.71 parts of secondary amine compound;

100-250 parts of diluent;

2-10 parts of disintegrating agent;

0-5 parts of glidant;

0-5 parts of lubricant;

0.01-15 parts of pharmaceutically acceptable acid;

preferably, the pharmaceutical composition comprises the following components in parts by weight:

1.71 parts of secondary amine compound;

184.47 parts of a diluent;

4 parts of disintegrating agent;

0.6 part of glidant;

5 parts of a lubricant;

1.71 to 3.42 portions of pharmaceutically acceptable acid

More preferably, the pharmaceutical composition is any one of the following formulations:

scheme one:

1.71 parts of valicarb isopropyl tartrate;

64 parts of anhydrous calcium hydrophosphate;

120.47 parts of microcrystalline cellulose;

4 parts of partially pregelatinized corn starch;

0.6 parts of colloidal silica;

5 parts of stearic acid;

3.42 parts of L- (+) -tartaric acid;

scheme II:

the pharmaceutical composition comprises the following components in parts by weight: 1.71 parts of valicarb isopropyl tartrate;

64 parts of anhydrous calcium hydrophosphate;

120.47 parts of microcrystalline cellulose;

4 parts of partially pregelatinized corn starch;

0.6 parts of colloidal silica;

5 parts of stearic acid;

1.71 parts of L- (+) -tartaric acid;

scheme III:

64 parts of anhydrous calcium hydrophosphate;

120.47 parts of microcrystalline cellulose;

4 parts of partially pregelatinized corn starch;

0.6 parts of colloidal silica;

5 parts of stearic acid;

3.42 parts of L- (+) -tartaric acid;

6 parts of Ophio-white or Ophio-blue;

scheme IV:

64 parts of anhydrous calcium hydrophosphate;

120.47 parts of microcrystalline cellulose;

4 parts of partially pregelatinized corn starch;

0.6 parts of colloidal silica;

5 parts of stearic acid;

1.71 parts of L- (+) -tartaric acid;

6 parts of Ophio-white or Ophio-blue.

9. The pharmaceutical composition according to claim 1 or 2, wherein the nitrosamine impurity content of the pharmaceutical composition is not greater than 7.4ppm, not greater than 7.3ppm, not greater than 7.2ppm, not greater than 7.1ppm, not greater than 7.0ppm; no more than 6.9ppm, no more than 6.8ppm, no more than 6.7ppm, no more than 6.6ppm, no more than 6.5ppm, no more than 6.4ppm, no more than 6.3ppm, no more than 6.2ppm, no more than 6.1ppm, no more than 6.0ppm; no more than 5.9ppm, no more than 5.8ppm, no more than 5.7ppm, no more than 5.6ppm, no more than 5.5ppm, no more than 5.4ppm, no more than 5.3ppm, no more than 5.2ppm, no more than 5.1ppm, no more than 5.0ppm; no more than 4.9ppm, no more than 4.8ppm, no more than 4.7ppm, no more than 4.6ppm, no more than 4.5ppm, no more than 4.4ppm, no more than 4.3ppm, no more than 4.2ppm, no more than 4.1ppm, no more than 4.0ppm, no more than 3.9ppm, no more than 3.8ppm, no more than 3.7ppm, no more than 3.6ppm, no more than 3.5ppm, no more than 3.4ppm, no more than 3.3ppm, no more than 3.2ppm, no more than 3.1ppm, no more than 3.0ppm; no more than 2.9ppm, no more than 2.8ppm, no more than 2.7ppm, no more than 2.6ppm, no more than 2.5ppm, no more than 2.4ppm, no more than 2.3ppm, no more than 2.2ppm, no more than 2.1ppm, or no more than 2.0ppm.

10. The preparation method according to any one of claims 1 to 9, characterized in that it comprises the following steps: mixing all the components, granulating, tabletting, and optionally coating;

preferably, the method comprises the steps of,

more preferably, the process is carried out,

the granulating is dry granulating or wet granulating;

the tabletting is a direct tabletting method.

11. Use of a pharmaceutical composition according to any one of claims 1 to 9 for the preparation of a medicament; the medicament is used for treating and/or preventing the following diseases: inflammatory bowel disease, ulcerative colitis, pyoderma gangrenosum, crohn's disease, irritable bowel syndrome, spasmodic dystonia, chronic pain, acute pain, diarrhea, cystitis, vasoconstriction, anxiety, mania, depression, bipolar disorder, autism, sleep disorder, jet lag, amyotrophic lateral sclerosis, cognitive dysfunction due to alcohol, barbiturates, vitamin deficiency, recreational drugs, lead, arsenic or mercury, alzheimer's disease, senile dementia, vascular dementia, parkinson's disease, multiple sclerosis, AIDS, encephalitis, trauma, hepatorenal encephalopathy, hypothyroidism, pick's disease, korskoff's syndrome, prefrontal dementia or subcortical dementia-induced cognitive dysfunction, hypertension, bulimia, anorexia, obesity, cardiac rhythm, diabetes mellitus, cerebral infarction, and other conditions Disorder, gastric acid hypersecretion, ulceration, pheochromocytoma, progressive supranuclear palsy, nicotine, tobacco products, alcohol, benzodiazepinesChemical dependence and addiction to barbiturates, opioids or cocaine, headache, migraine, stroke, traumatic brain injury, obsessive-compulsive disorder, psychosis, huntington's chorea, tardive dyskinesia, hyperkinesia, dyskinesia, schizophrenia, multi-infarct dementia, age-related cognitive decline, epilepsy, attention deficit hyperactivity disorder or tourette's syndrome;

preferably, the medicament meets one of the following conditions a to c:

c. the medicine is a smoking cessation medicine.

12. Use of a pharmaceutically acceptable acid to inhibit or slow down nitrosamine amination of a secondary amine compound; wherein the secondary amine compound is valicarb or a pharmaceutically acceptable salt thereof;

preferably, the use of said pharmaceutically acceptable acid in inhibiting or slowing down the nitrosation of a secondary amine compound satisfies one or more of the following conditions la to id:

The molar ratio of la, the secondary amine compound and the pharmaceutically acceptable acid is 1 (0.01-50), e.g., a molar ratio of 1:0.05, 1:0.1, 1:0.5, 1:1, 1:1.5, 1:2, 1:2.5, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, or 1:40;

id. The pharmaceutically acceptable acid is added in an amount such that the pH value of the mixture X is less than or equal to 4 under the following test conditions; the mixture X comprises the pharmaceutically acceptable acid in admixture with the secondary amine compound, preferably at a pH of 1 to 4, for example 1, 1.5, 2, 2.5, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9 or 4, etc.; when the mixture is solid, the test conditions are: mixing the mixture with water with the pH value of 6+/-0.1 to form a dispersion liquid with the concentration of 20%, and testing the obtained pH value; when the mixture is liquid, the test conditions are: the solvent of the liquid is water with pH=6+/-0.1, the concentration of the liquid is 20%, and the pH value obtained by testing is measured;

More preferably, the use of said pharmaceutically acceptable acid for inhibiting or slowing down the nitrosation of a secondary amine compound satisfies one or more of the following conditions IIa to IIc:

IIc, wherein the pharmaceutically acceptable acid is a solid acid; further preferably, the pharmaceutically acceptable acid is tartaric acid, for example, L- (+) -tartaric acid or D- (-) -tartaric acid;

it is further preferred that the composition comprises,

the application of the pharmaceutically acceptable acid in inhibiting or slowing down the nitrosamine of the secondary amine compound simultaneously meets the conditions Ia-ic or IIa-IIc.

13. A method for inhibiting or slowing down the nitrosamine of a secondary amine compound, characterized in that a secondary amine compound or a composition thereof is mixed with a pharmaceutically acceptable acid; wherein the secondary amine compound is valicarb or a pharmaceutically acceptable salt thereof;

preferably, the method for inhibiting or slowing down the nitrosamine of the secondary amine compound comprises the steps of: adding pharmaceutically acceptable acid into the secondary amine compound or the composition thereof, and uniformly mixing;

Preferably, the composition of secondary amine compounds comprises all components of the pharmaceutical composition of any one of claims 1 to 9 except pharmaceutically acceptable acids;

preferably, the method for inhibiting or slowing down the nitrosamine of the secondary amine compound satisfies one or more of the following conditions ia to id:

the molar ratio of the Ia to the secondary amine compound to the pharmaceutically acceptable acid is 1:0.01-50; for example, the molar ratio is 1:0.05, 1:0.1, 1:0.5, 1:1, 1:1.5, 1:2, 1:2.5, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, or 1:40;

more preferably, the method for inhibiting or slowing down the nitrosamine of the secondary amine compound satisfies one or more of the following conditions IIa to IIc:

IIc, wherein the pharmaceutically acceptable acid is a solid acid; further preferably, the pharmaceutically acceptable acid is tartaric acid, e.g., L- (+) -tartaric acid or D- (-) -tartaric acid;

It is further preferred that the composition comprises,