Patent Description:
Expectoration is a common symptom of respiratory diseases. The increase of sputum can stimulate the respiratory mucosa and cause cough. Blocking of the bronchioles can not only cause asthma, but also cause secondary infection, further damages the respiratory tract, and aggravates cough, expectoration and asthma. In severe cases, breathing may be suppressed or suffocation may be resulted. Excessive secretion of mucus can cause dysfunction of mucociliary clearance and damage to local defense function, resulting in uncontrollable infection and in aggravation of airway obstruction, which directly affects the progression of the disease and the subjective feelings of patients. Therefore, the use of expectorants to promote the efflux of airway secretions as soon as possible is an important auxiliary measure for the treatment of airway inflammation.

Chinese patent application <CIT> discloses an ambroxol derivative and a method of preparing this derivative. This derivative can be used in medical treatment Another Chinese patent application <CIT> discloses an ambroxol derivative, a preparation method therefor, and this ambroxol derivative in drugs for eliminating phlegm.

Expectorants are a class of drugs that can thin sputum, reduce its viscosity to facilitate expectoration of sputum or can accelerate the movement of mucociliary in the respiratory tract and improve the function of sputum transport. Expectorants promote the discharge of build-up phlegm in the lumen of the respiratory tract, reduce the irritation to the respiratory mucosa, play the role of antitussive and asthma indirectly, and also help control secondary infections. Expectorant drugs play the role of promoting phlegm efflux in the following aspects: (<NUM>) improve the physical and chemical properties of sputum and reduce viscosity. (<NUM>) restore the normal structure of airway epithelial mucus layer and the function of ciliary clearance. (<NUM>) inhibit the production and secretion of mucin, and reduce the production of high-viscosity mucus. (<NUM>) promote DNA depolymerization in purulent sputum. (<NUM>) resist inflammation and reduce DNA production. Based on the main effect of the drug, expectorants are divided into mucus secretion promoting drugs (nausea expectorants and irritating expectorants) and mucolytics. Currently, clinically representative chemical expectorants include ammonium chloride, acetylcysteine, ambroxol, erdosteine, etc. However, the above commonly used drugs all have certain adverse reactions, thus it is necessary to develop a new expectorant that is safe and effective.

The present disclosure relates to compounds represented by formula I and formula II and their non-toxic pharmaceutically acceptable salts, as well as pharmaceutical compositions containing these compounds as active ingredient, and the use of the compounds and pharmaceutical compositions in expectorants. In the present disclosure, by introducing the sulfhydryl group in acetylcystine of the compound into ambroxol or similar structure, the inventors surprisingly found that the therapeutic effect of expectorant is significantly better than that of the existing expectorant. The compounds of the present disclosure have higher safety and improved therapeutic effect.

The first aspect of the present disclosure therefore provides compounds represented by formula I and formula II, and pharmaceutically acceptable salts thereof:
<CHM>
wherein in the compound represented by formula I:.

Preferably, the present disclosure provides compounds represented by formula I and formula II, or pharmaceutically acceptable salts thereof, and the compound is selected from the compounds represented by the following formulas:.

Further, described herein is a method for preparing the compounds represented by the formula I and formula II, which are prepared through the following steps;
<CHM>
<CHM>
<CHM>.

The second aspect of the present disclosure relates to a pharmaceutical composition comprising at least one compound represented by formula I and formula II, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers or excipients.

The third aspect of the present disclosure relates to compounds of formula I and formula II and non-toxic pharmaceutically acceptable salts thereof, and to the use of compounds of formula I and formula II and non-toxic pharmaceutically acceptable salts thereof as active ingredient in the pharmaceutical composition as an expectorant.

The compounds represented by formula I and formula II can form pharmaceutically acceptable salts with inorganic acids such as hydrochloride, hydrobromide, sulfate and the like. Selection and preparation of appropriate salts are well known to those skilled in the art.

The compounds of the present disclosure, or pharmaceutically acceptable salts thereof, may be administered alone or in a form of pharmaceutical compositions. The pharmaceutical compositions of the present disclosure can be formulated into various suitable dosage forms based on the administration route. One or more physiologically acceptable carriers including excipients and auxiliaries are employed, which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Appropriate forms of formulation depend on the route of administration chosen and can be prepared according to general knowledge well known in the art.

The route of administration can be oral, parenteral or topical, preferably oral, aerosol inhalation and injection. Pharmaceutical preparations that can be administered orally include oral liquids, granules or tablets and the like. Those skilled in the art will understand that the compounds of the present disclosure can be used with a suitable drug delivery system (DDS) to produce more favorable effects.

The following examples facilitates comprehensive understanding of the present disclosure, however, they do not limit to the present disclosure in any way.

To a <NUM> reaction flask were added <NUM> of trans-<NUM>-acetamidocyclohexanethiol, <NUM> of potassium hydroxide and <NUM> of water, and the reaction flask was placed in an oil bath, and heated under reflux for <NUM> hours. Upon termination of the reaction, the reaction mixture was extracted with dichloromethane (<NUM>×<NUM>); the organic phases were combined, dried over anhydrous sodium carbonate, filtered. Hydrogen chloride was introduced into the filtrate, and when no solid were further produced, the filtrate was cooled down to <NUM> and was stirred and crystallized for <NUM> hours, and filtered to give <NUM> of trans-<NUM>-aminocyclohexanethiol hydrochloride.

To a <NUM>-ml three-necked bottle were added trans-<NUM>-aminocyclohexanethiol hydrochloride (<NUM>), sodium hydroxide (<NUM>), potassium carbonate (<NUM>), and ethylene glycol monomethyl ether (<NUM>), and the bottle was place in an oil bath, and heated to <NUM>. N-(<NUM>-amino-<NUM>,<NUM>-dibromo-benzoyl)-N-methanesulfonyl hydrazide (<NUM>) was added in <NUM> batched. Upon completion of addition, the resulting mixture was heated under reflux for <NUM>, and then the reaction system was cooled to room temperature. Water (<NUM>) was added and stirred for crystallization. The crystals were filter to afford <NUM> of trans-<NUM>-[(<NUM>-amino-<NUM>,<NUM>-dibromo-benzylidene)amino]-cyclohexanethiol.

To a hydrogenation reactor were added trans-<NUM>-[(<NUM>-amino-<NUM>,<NUM>-dibromo-benzylid-ene)amino]-cyclohexanethiol (<NUM>), and glacial acetic acid <NUM>, 5wt% pd/C <NUM>. And hydrogenation was carried out for <NUM> at <NUM>. 2MPa, <NUM>. Upon completion of the reaction, the solvents were removed under reduced pressure, and the resulting residue was dissolved in <NUM> of acetone, then <NUM> of hydrochloric acid was added dropwise. The temperature was lowered to <NUM> and crystallized for <NUM>, the crystals were filtered and dried to afford trans-<NUM>-[(<NUM>-amino-<NUM>,<NUM>-dibromo-benzyl)amino]-cyclohexanethiol hydrochloride (I1).

The operations were referred to Examples <NUM>, <NUM>, and <NUM>, except that trans-<NUM>-aminocyclohexanethiol hydrochloride is replaced with <NUM>-amino cyclohexanethiol hydrochloride to give the following compound of formula I.

The operations were referred to Examples <NUM>, <NUM>, and <NUM>, except that trans-<NUM>-aminocyclohexanethiol hydrochloride is replaced with <NUM>-methylamino-cyclohexanethiol hydrochloride to afford compound I6.

The operations were referred to Example <NUM>, except that <NUM>-methylamino-cyclohexanethiol hydrochloride was replaced with methylamino-cyclohexanethiols having different -SH group positions and isomers to give the compound of formula I below.

Ten grams of cis-<NUM>-(<NUM>-chloro-<NUM>-nitro-benzylamino)-cyclohexanethiol hydrochloride was dissolved in <NUM> of ethanol and <NUM> of water, the resulting solution was mixed with <NUM> of palladium carbon, and hydrogenation was carried out at room temperature to reduce the resulting mixture. After the hydrogen absorption is completed, the palladium carbon was removed by filtration, and a small amount of ether was added to the filtrate to precipitate a solid, and the solids were collected and recrystallized from ethanol to give the compound (I9).

Thirty grams of lithium aluminum hydride was added to <NUM> of anhydrous tetrahydrofuran, and the mixture was added dropwise to a solution of <NUM> of cis-<NUM>-amino-<NUM>-chloro-N-(<NUM>-mercapto-cyclohexyl)-benzamide in anhydrous tetrahydrofuran. After all the mixture was dripped, the resulting mixture was and refluxed with stirring for <NUM> hours. Then ethyl acetate and 5N sodium hydroxide were added to quench the unreacted lithium aluminum hydride. Filtration was conducted and the filtrate was concentrated to dryness under vacuum. The residue was purified by silica gel column chromatography to give compound (I10).

The operations were referred to Example <NUM>, the difference was that different benzamide compounds were reduced to afford the following compounds of formula I.

Compound I1 (<NUM>, <NUM>. 057mol) was added into <NUM> of water, stirred to dissolve all the solids, and <NUM>. <NUM> (<NUM>. 172mol) allyl alcohol, <NUM> (<NUM>. 0037mol) K<NUM>S<NUM>O<NUM>, and <NUM> (<NUM>. 0037mol) NaHSO<NUM> were added in sequence under stirring. The resulting mixture was heated to <NUM>, stirred for <NUM> hours. After the reaction was completed, water and excess allyl alcohol were distilled off, <NUM> of absolute ethanol was added, stirred to precipitate a solid, and the solids were purified to give Compound I13.

In this example, the in vivo efficacy of the compounds I1, I4, I13 of the present disclosure and known compounds was tested.

Ninety healthy mice were randomly divided into <NUM> groups, and the specific grouping is shown in Table <NUM>. Mice in the positive control group were give medicinal ammonium chloride and ambroxol hydrochloride orally, mice in the normal control group were given normal saline of the same amount, and mice the other groups were given corresponding drugs by gavage. The mice were administered by gavage once in the morning and afternoon (excluding ammonium chloride) <NUM> days before the experiment. After <NUM> hour of intragastric administration on the morning of the experiment, the mice were intraperitoneally injected with <NUM> of phenol red, and the mice were sacrificed <NUM> hours later. The tissue surrounding the organs was peeled off, and a section of the trachea from the thyroid cartilage to the branch of the trachea was cut off and placed in a test tube containing <NUM> of normal saline. The tube was shaken for <NUM> minutes, the trachea was discarded, and <NUM> of sodium hydroxide (<NUM> mol/L) was added to each sample solution. The OD value was measured using a UV-visible spectrophotometer at a wavelength of <NUM>, and the absorbance values were converted into the phenol red content according to the phenol red standard curve.

The data was processed by PEMS medical statistical software. The results showed that each medication group significantly promoted the secretion of phenol red in the tracheal segment of mice. The ambroxol group and the high dose of compounds I1 and I13 had the best effect, and the compound I13 had the strongest effect. The effect produced by each dose was significantly better than that of ammonium chloride and ambroxol, among which compound I13 had the most significant expectorant effect. It is suggested that the expectorant effect of compound I13 is enhanced with the increase of dose, and there is a relatively obvious dose-effect relationship, and compound I13 has better the effect than compound I1.

To test the acute toxicity of compounds of the present disclosure and comparative compounds, the following experiments were performed.

Compounds of the present disclosure were dissolved in water and administered to <NUM> ICR mice (mice of <NUM>-week old, male, weighing <NUM> ± <NUM>). Intravenous administration was performed to determine the median lethal dose (LD50, mg/kg). Ambroxol hydrochloride was used as a control. The results are shown in the table below.

The test results show that the LD50 values for compounds I1, I4 and I13 are much higher than that for ambroxol hydrochloride, suggesting that the safety of the compounds of the present disclosure is better than that of ambroxol hydrochloride.

Formula used in the Examples: preparation of pharmaceutical composition.

The above materials were mixed, and the mixture was filled into a sealed package to prepare a powder. Preparation of tablets.

The above materials were mixed and the mixture was then compressed into tablets by known methods. Preparation of capsules.

Capsules were prepared by mixing the above materials and filling the mixture into gelatin capsules by known methods. Preparation of injections.

The compound of Example <NUM> and glucose were dissolved in water, the pH was adjusted to <NUM>-<NUM> with a pH adjuster, and the solution was freeze-dried in a freeze-drying oven. After drying, the vials were plugged and capped. Preparation of inhalants.

Claim 1:
A compound represented by formula I, pharmaceutically acceptable salt or tautomer thereof;
<CHM>
wherein R<NUM> represents H, F, Cl, Br or NH<NUM>; R<NUM> represents H, F, Cl or Br; R<NUM> represents H, F, Cl, Br or NH<NUM>; R<NUM> represents H, F, Cl or Br; R<NUM> represents H, F, Cl or Br; R<NUM>
represents H or CH<NUM>; and R<NUM> represents
<CHM>
wherein R<NUM> is not
<CHM>
if:
R<NUM>, R<NUM>, R<NUM>, R<NUM>, and R<NUM> are all H, and
the compound represented by formula I is not a pharmaceutically acceptable salt thereof.