WO2022007743A1 - Mucosal administration dosage form and use thereof - Google Patents

Abstract

Provided is a mucosal administration dosage form, in particular a nasal administration dosage form, containing a compound of formula I or a pharmaceutically acceptable salt, a solvate, or a prodrug thereof. The mucosal administration dosage form provided in the present invention can be used to treat CCR4-mediated diseases, particularly asthma, COPD and rhinitis. The nasal mucosal administration dosage form provided in the present invention has the advantages of being quick-acting, avoiding the first-pass effect and being high in bioavailability, and is convenient to use, safe and good in terms of compliance.

Description

Forms for mucosal administration and their uses

This application claims the priority of the Chinese patent application filed on July 7, 2020 with the application number 202010646239.1 and the invention titled "Nasal mucosal administration dosage form and its application", the entire contents of which are incorporated into this application by reference.

Field of Invention

The invention belongs to the field of medicine, and relates to a mucosal administration dosage form containing a CCR4 inhibitor, and the use of the medicine for treating diseases mediated by CCR4.

Background technique

CCR4 (Chemokine Receptor 4, Chemokine Receptor 4) was first discovered by Christine A.Power in 1995 (Christine AP et al. J.Biol.Chem.1995,270(8):19495-19500), belonging to chemotaxis A member of the factor receptor (CCR) family, it is a seven-transmembrane G-protein coupled receptor. CCR4 is expressed in peripheral blood leukocytes, thymocytes, basophils, monocytes, macrophages, platelets, IL-activated NK cells, spleen and brain, and can play an important role in a variety of diseases. For example, when human allergic dermatitis (AD) occurs, the expression of CCR4 expressed by CD4+ T cells increases in peripheral blood mononuclear cells (PBMCs), and the level of TARC in serum also increases accordingly. This suggests that the chemotaxis of CCR4 expression in cells is induced by TARC and selectively induces Th2 cell migration towards damaged skin in human atopic dermatitis. The drugs used to treat allergic dermatitis mainly include antihistamines and bronchodilators, but they can only improve symptoms, but have no effect on the development of the disease. In addition, corticosteroids also have a certain effect on allergic dermatitis, but there are potential safety hazards. Studies have shown that antagonism of MDC or TARC can reduce the accumulation of T cells at the site of inflammation, and CCR4 antagonists may be very effective in the treatment of allergic inflammation including but not limited to allergic rhinitis and allergic dermatitis.

CCR4 has also been found to be strongly associated with lung diseases such as chronic obstructive pneumonia, chronic bronchitis and asthma. CCR4 can be restrictedly expressed in the cells involved in the asthma response, and is considered to be a good target for the treatment of asthma. Therefore, the development of CCR4 antagonist drugs has good application prospects.

CN101370793A discloses compounds of formula and their use as CCR4 antagonists, including for CCR4-mediated diseases

wherein each substituent is as defined as disclosed in this application.

CN102388039A discloses indazole compounds of the following formula (I) and their use as CCR4 antagonists, including their use in treatments associated with CCR4 antagonists:

wherein each substituent is as defined as disclosed in this application.

Anthony Cahn et al. disclosed the phase I clinical results of the compound GSK2239633 in the above structure (Cahn et al. BMC Pharmacology and Toxicology 2013, 14: 14), the experiment showed that the compound has good human tolerance, but in the maximum safety The body exposure at the dose is too low to produce the expected medicinal effect.

There remains a need in the art for better CCR4 inhibitor drugs or dosage forms.

SUMMARY OF THE INVENTION

The invention provides a technology for improving the bioavailability of GSK2239633 through mucosal administration route and a usable pharmaceutical preparation, which solves the problems of low solubility of CCR4 inhibitors in the prior art, difficulty in preparation, and excessive bioavailability of the prepared preparations. Low, unable to reach the expected effective plasma concentration in the body. The CCR4 inhibitor pharmaceutical preparation provided by the present invention also has the advantages of no local irritation and safe use.

Specifically, the present invention provides a dosage form for mucosal administration, which contains a compound of formula I or a pharmaceutically acceptable salt, solvate or prodrug thereof:

The CAS number of the compound of formula I is 1240516-71-5, its preparation is disclosed in Example 57 of CN102388039A, and its chemical name is N-[(3-{[3-{[(5-chloro-2-thienyl) Sulfonyl]amino}-4-(methoxy)-1H-indazol-1-yl]methyl}phenyl)methyl]-2-hydroxy-2-methylpropanamide. In the art, the common name for the compound of formula I is GSK2239633.

The compounds of formula I may be administered in the form of prodrugs, which are compounds that are cleaved in humans or animals to release the compounds of the present invention. Examples of prodrugs include in vivo cleavable ester derivatives and in vivo cleavable amide derivatives, which may be formed at the carboxyl or hydroxyl group of the compound of formula I, which amide derivatives may be formed at the carboxyl group of the compound of formula I or on the amino group.

Pharmaceutically acceptable prodrugs of the compounds of formula I are, for example, their in vivo cleavable esters or ethers. In vivo cleavable esters or ethers of hydroxyl-containing compounds of formula I are, for example, pharmaceutically acceptable esters or ethers that are cleaved in humans and animals to yield the parent hydroxy compound. For hydroxyl groups, suitable pharmaceutically acceptable ester-forming groups include inorganic esters such as phosphates (including phosphoramidates). For hydroxyl, suitable pharmaceutically acceptable ester-forming groups also include (1-10C) alkanoyl groups (such as acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl), (1-10C) alkanoyl groups 10C) Alkoxycarbonyl (eg ethoxycarbonyl, N,N-[di-(1-4C)alkyl]carbamoyl, 2-dialkylaminoacetyl and 2-carboxyacetyl). Examples of ring substituents on phenylacetyl and benzoyl include aminomethyl, N-alkylaminomethyl, N,N-dialkylaminomethyl, morpholinomethyl, piperazin-1-ylmethyl and 4-(1-4C)alkylpiperazin-1-ylmethyl. For hydroxyl, suitable pharmaceutically acceptable ether-forming groups include alpha-acyloxyalkyl groups such as acetoxymethyl and pivaloyloxymethyl.

The compounds of formula I of the present invention, or pharmaceutically acceptable salts, solvates or prodrugs thereof, have CCR4 inhibitory activity. Thus, a compound of formula I or a pharmaceutically acceptable salt, solvate or prodrug thereof can be used for the treatment of CCR4 mediated diseases or for the manufacture of a medicament for the treatment of CCR4 mediated diseases.

In yet another aspect of the invention, the dosage form for mucosal administration is a dosage form for nasal administration. In yet another aspect of the present invention, the dosage form for mucosal administration is nasal administration and is a solution composition containing the compound of formula I or a pharmaceutically acceptable salt, solvate or prodrug thereof. Dosage forms for nasal administration can be administered by inhalation of the composition using a suitable delivery device such as a nebulizer (eg, a spray pump, aerosol, nebulizer, or metered-dose nebulizer) or a dropper container (eg, a dropper or nasal dropper). It is dropped or applied in the nasal cavity to adhere to the nasal mucosa, whereby the active ingredient is absorbed through the nasal mucosa. In the present invention, the compound of formula I above is substantially completely dissolved in the solution. In certain instances, the dosage form for mucosal administration is in the presence of a partially suspended compound of formula I.

In yet another aspect of the invention, the dosage form for mucosal administration, particularly a dosage form for transnasal mucosal administration, eg, a solution for spraying, comprises a cosolvent. In the present invention, the cosolvent facilitates the dissolution of the compound of formula I in solution. In one aspect of the present invention, the cosolvent is selected from sodium hydroxide, sodium methoxide, potassium hydroxide, sodium bicarbonate, sodium dihydrogen phosphate, sodium citrate, α-, β- or γ-cyclodextrin any one or more of them. In yet another aspect of the present invention, the co-solvent is sodium hydroxide and/or β-cyclodextrin. In yet another aspect of the present invention, the co-solvent is sodium hydroxide.

In yet another aspect of the present invention, the content of the cosolvent contained in the mucosal administration dosage form is about 0.25-1.5 mol/L, preferably 0.5-1.0 mol/L.

In yet another aspect of the invention, the dosage form for mucosal administration, particularly a dosage form for transnasal mucosal administration, eg, a solution for spraying, comprises a pH adjusting agent. In yet another aspect of the present invention, the mucosal administration dosage form comprises a pH adjusting agent selected from the group consisting of phosphoric acid, hydrochloric acid, citric acid, tartaric acid, acetic acid, maleic acid, sodium hydroxide, sodium carbonate, sodium bicarbonate, citric acid Any one or more of sodium, phosphate buffer, acetate buffer, and citrate buffer. Preferably, the dosage form for mucosal administration comprises a pH adjusting agent phosphate and/or citrate buffer.

In yet another aspect of the present invention, the dosage form for mucosal administration, particularly a dosage form for transnasal mucosal administration, eg, a solution for spraying, comprises a cosolvent and a pH adjuster.

In one aspect of the invention, the mucosal administration dosage form has a pH of about 6.5-9.5; preferably about 7.0-8.5, such as 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9 , 8.0, 8.1, 8.2, 8.3, 8.4, or 8.5, and a range of pH values formed by any two of these specific pH values.

Without being bound by theory, when the pH is higher than 9.5, the irritation to the nasal cavity is greater, although the solubility of the compound of formula I is ideal at this time. The inventors have also found in research that when the pH is low, for example, at or below 6.5, the compound of formula I is easily precipitated.

In yet another aspect of the invention, the mucosal administration dosage form comprises a preservative, for example selected from methylparaben, ethylparaben, propylparaben, butylparaben, Sodium Methylparaben, Sodium Ethylparaben, Sodium Propylparaben, Sodium Butylparaben, Sorbic Acid, Sodium Sorbate, Benzoic Acid, Sodium Benzoate, Benzyl Alcohol, Benzalkonium Any one or more of ammonium bromide, benzalkonium chloride, chlorobutanol, resorcinol and sodium edetate, preferably, the preservative is sodium methylparaben.

In yet another aspect of the invention, the dosage form for mucosal administration is an isotonic formulation. Since the nasal environment is capable of self-regulating, in yet another aspect of the present invention, the formulation is free of osmotic pressure regulators.

In yet another aspect of the present invention, the mucosal administration dosage form comprises an osmotic pressure regulator, such as any one or more selected from sodium chloride, potassium nitrate, boric acid and glucose, preferably sodium chloride.

In yet another aspect of the invention, the mucosal administration dosage form does not contain a penetration enhancer. The compound of formula I has a molecular weight of about 550, and the formulation may be free of penetration enhancers.

In one aspect of the present invention, the mucosal administration dosage form spray solution dosage form has a spray particle size of about 5-200 μm, preferably about 10-150 μm.

In one aspect of the present invention, the dosage form for mucosal administration (especially the dosage form for transnasal mucosal administration, such as a solution for spraying) is present in an amount of the compound of formula I or a pharmaceutically acceptable form thereof per 100 ml of the dosage form. The amount of salt, solvate or prodrug is about

1.25 mg-10.0 g, preferably about 2.5 mg-5 g, and preferably about 5 mg-1 g.

In one aspect of the invention, the dosage form for mucosal administration (especially the dosage form for transnasal mucosal administration, eg, a solution for spraying) is a dosage form for single-dose or multiple-dose administration, wherein the The amount of the compound of formula I, or a pharmaceutically acceptable salt, solvate or prodrug thereof, is about 2.5 μg to 10.0 mg, preferably about 5.0 μg to 1.0 mg.

In one aspect of the invention, the mucosal administration dosage form is used to treat a CCR4 mediated disease. A compound of formula I, or a pharmaceutically acceptable salt, solvate or prodrug thereof, has CCR4 inhibitory activity. Thus, the compounds of formula I of the present invention or pharmaceutically acceptable salts, solvates or prodrugs thereof can be used for the treatment of CCR4 mediated diseases or for the preparation of medicaments for the treatment of CCR4 mediated diseases. In particular, the compounds of formula I of the present invention, or pharmaceutically acceptable salts, solvates or prodrugs thereof, can be used to treat diseases in which modulation of CCR4 (eg, inhibition of CCR4 activity) is beneficial to patients.

The CCR4-mediated diseases are, for example: (1) respiratory diseases, such as airway obstructive diseases, including asthma, including bronchial asthma, allergic asthma, endogenous asthma, extrinsic asthma, exercise-induced asthma, Drug-induced (including aspirin and NSAID-induced) asthma and dust-induced asthma, intermittent and persistent asthma; chronic obstructive pulmonary disease (COPD); bronchitis, including infectious and eosinophilic bronchitis; pulmonary gas Bronchiectasis; cystic fibrosis; hypersensitivity pneumonitis; pulmonary fibrosis, including cryptogenic fibrotic alveolitis, idiopathic interstitial pneumonia, fibrosis complicated by antineoplastic therapy and chronic infection; pulmonary vascular Vasculitis and thrombotic disease and pulmonary hypertension; rhinitis: acute and chronic rhinitis, including drug-induced rhinitis and vasomotor rhinitis; perennial and seasonal allergic rhinitis, including neuropathic rhinitis; acute viral infection , including the common cold and infections caused by respiratory syncytial virus, influenza, coronavirus and adenovirus. Preferably, wherein the CCR4 mediated diseases are asthma, COPD and rhinitis.

In one aspect of the present invention there is provided a compound of formula I or a pharmaceutically acceptable salt, solvate or prodrug thereof for use in a method for the treatment or prevention of CCR4 mediated diseases by mucosal route of administration, in particular Methods of treating or preventing CCR4-mediated diseases by the nasal mucosal route of administration. In the method, the compound of formula I, or a pharmaceutically acceptable salt, solvate or prodrug thereof, is administered in the form of a spray solution.

According to yet another aspect of the present invention there is provided a compound of formula I or a pharmaceutically acceptable salt, solvate or prodrug thereof in the manufacture of a medicament for the treatment or prevention of CCR4 mediated diseases by mucosal route of administration Use, in particular by the nasal mucosal route of administration, eg in the form of a spray solution.

The present invention also provides a method of preparing a dosage form for mucosal administration of the present invention as previously defined, wherein the mucosal dosage form comprises a compound of formula I or a pharmaceutically acceptable salt, solvate or prodrug thereof.

In one aspect of the invention, the dosage form for mucosal administration is a dosage form for nasal administration, eg, a solution composition. In the present invention, the method includes the step of mixing a compound of formula I, or a pharmaceutically acceptable salt, solvate or prodrug thereof, with a co-solvent and/or pH adjusting agent.

In one aspect of the invention, the method includes the step of adjusting the pH to about 6.5-9.5; preferably about 7.0-8.5. Optionally, the method further includes the step of adding one or more of preservatives, osmotic pressure regulators, and penetration enhancers. Wherein, the cosolvent, pH adjuster, preservative, osmotic pressure adjuster, and penetration enhancer are as defined above.

In one of the embodiments of the present invention, the method for preparing the nasal dosage form of the present invention, especially the spray, comprises the following steps:

1. Dissolving a compound of formula I or a pharmaceutically acceptable salt, solvate or prodrug thereof using a cosolvent, adding a pH adjuster to adjust the pH of the solution to about 7.0-9.0, for example to about 8.0,

The volume of the cosolvent added therein is about 0.5-3 parts by volume, preferably about 1-2 parts by volume, for example, about 1.5 parts by volume;

2. Add water and/or other components to the volume of the intended formulation, and adjust the pH to about 6.5-9.5; preferably about 7.0-8.5.

In one aspect of the present invention, the cosolvent added in step 1 of the preparation method is selected from sodium hydroxide, sodium methoxide, potassium hydroxide, sodium bicarbonate, sodium dihydrogen phosphate, sodium citrate, α-, β - or any one or more of γ-cyclodextrin, preferably, the co-solvent it contains is sodium hydroxide/or β-cyclodextrin, more preferably, the co-solvent it contains is sodium hydroxide and .

In one aspect of the invention, the formulation for mucosal administration of the compound of formula I is prepared by first dissolving the compound of formula I in a smaller volume of co-solvent (eg, sodium hydroxide), and then adding the other components or water to the working volume. In yet another aspect of the present invention, the concentration of the cosolvent added in the mucosal administration dosage form is about 0.1-3.0 mol/L, preferably about 0.2-2.0 mol/L, more preferably about 0.3-1.0 mol/L L, for example, is about 0.5 mol/L, and contains about 0.5-3.0 parts by volume, preferably about 1.0-2.0 parts by volume, such as about 1.5 parts by volume, of cosolvent per 100 parts by volume of the dosage form for mucosal administration.

The inventors found in their research that co-solvents such as sodium hydroxide can promote the dissolution of the compound of formula I, but when the amount of sodium hydroxide is low or the concentration of sodium hydroxide in the formulation is low, it is not enough to dissolve the CCR4 inhibitor . When the dosage of sodium hydroxide is higher or the concentration of sodium hydroxide in the formulation is higher, more pH adjusters such as sodium dihydrogen phosphate and sodium bicarbonate need to be used. The inventors found that in this case, the osmotic pressure of the prepared preparation was high, which was unfavorable for nasal administration; and the inventors also found that in this case, a large amount of heat would be generated, which was unfavorable for the preparation process In particular, it is not conducive to industrial production. Therefore, in one of the embodiments, in preparing the nasal spray containing the CCR4 inhibitor of formula 1 per 100ml, the amount of sodium hydroxide added is calculated at a concentration of about 0.5mol/L, and the volume is about 0.5ml-3ml , preferably about 1ml-2.0ml, more preferably 1.5ml. Other concentrations of sodium hydroxide may also be used so long as the amount of sodium hydroxide falls within or is equivalent to the above ranges. In one of the embodiments, it is preferred to use sodium hydroxide with a higher concentration, such as sodium hydroxide greater than or equal to 0.1 mol/L, preferably greater than 0.3 mol/L sodium hydroxide; more preferably, the concentration is greater than or equal to 0.1 mol/L. 0.5mol/L sodium hydroxide.

The methods and formulations provided by the present invention have but are not limited to the following advantages:

1. The compound of formula I has good solubility and high solubility in the formulation of the present invention, the nasal ciliary toxicity is small and reversible, and the pH value of the formulation is suitable for the nasal cavity.

2. Compared with the conventional route of administration, the nasal spray provided by the present invention has the advantages of quick-acting, avoiding the first-pass effect, high bioavailability, convenient use, safety and good compliance.

detailed description

The substance and beneficial effects of the present invention will be further described below with reference to the embodiments. The examples are only used to illustrate the present invention and not to limit the present invention.

GSK2239633, CAS number 1240516-71-5, was purchased from MCE China, catalog number: HY-100183. The reagents or instruments used in the examples without the manufacturer's indication are conventional products that can be obtained from the market. If the specific conditions are not indicated in the examples, it is carried out according to the conventional conditions or the conditions suggested by the manufacturer.

Example 1: Preparation of CCR4 inhibitor nasal spray 1

Weigh 0.03g of sodium methylparaben, put it in a 100ml measuring bottle, add an appropriate amount of water for injection to dissolve, then add 250mg of GSK2239633, add an appropriate amount of sodium hydroxide (1.5ml, 0.5mol/L), shake to make Dissolve, add water for injection to the scale of 100ml, shake well, adjust the pH to 7.0 with sodium dihydrogen phosphate, add 0.3g of sodium chloride, add water for injection to the mark, shake well, filter the medicinal solution with a 0.22μm microporous membrane , the nasal spray liquid of CCR4 inhibitor was obtained, and the pH was about 7.5. It is then filled into single-dose and multi-dose nasal spray devices, respectively, and packaged for later use.

Example 2: Preparation of CCR4 inhibitor nasal spray 2

Weigh 0.03g of sodium methylparaben, put it in a 100ml measuring bottle, add an appropriate amount of water for injection to dissolve, then add 250mg of GSK2239633, add an appropriate amount of sodium hydroxide (1.5ml, 0.5mol/L), shake to make Dissolve, add 15.0g of propylene glycol, add water for injection to the mark of 100ml, shake well, adjust the pH value to 8.0 with sodium bicarbonate, add 0.3g of sodium chloride, add water for injection to the mark, shake well, use 0.22μm micropore The medicinal solution is filtered through a filter membrane to obtain a nasal spray medicinal solution of a CCR4 inhibitor with a pH of about 8.5. It is then filled into single-dose and multi-dose nasal spray devices, respectively, and packaged for later use.

Example 3: Preparation of CCR4 inhibitor nasal spray 3

Weigh 0.03g of sodium methylparaben, put it in a 100ml measuring bottle, add an appropriate amount of water for injection to dissolve, then add 250mg of GSK2239633, add an appropriate amount of sodium hydroxide (1.5ml, 0.5mol/L), shake to make Dissolve, add 15.0g of propylene glycol, add water for injection close to the mark, shake well, add citric acid and sodium citrate (1:10), adjust the pH value to 8.0, add water for injection to the mark of 100ml, shake well, use a 0.22μm micrometer The medicinal solution is filtered through a pore filter to obtain a nasal spray medicinal solution of a CCR4 inhibitor with a pH of about 8.5. It is then filled into single-dose and multi-dose nasal spray devices, respectively, and packaged for later use.

Example 4: Preparation of CCR4 inhibitor nasal spray 4

Weigh 0.03g of sodium methylparaben, put it in a 100ml measuring bottle, add an appropriate amount of water for injection to dissolve, then add 1.0g of GSK2239633, add an appropriate amount of sodium hydroxide (1.5ml, 0.5mol/L), shake To dissolve, add water for injection close to the mark, shake well, adjust the pH value to 7.0 with sodium hydrogen phosphate, add 0.3 g of sodium chloride, add 2.5 g of β-cyclodextrin, add water for injection to the mark, shake well, use 0.22 The drug solution was filtered through a μm microporous membrane to obtain a nasal spray drug solution of a CCR4 inhibitor with a pH of about 7.5. It is then filled into single-dose and multi-dose nasal spray devices, respectively, and packaged for later use.

Example 5: Preparation of CCR4 inhibitor nasal spray 5

Weigh 0.03g of sodium methylparaben, put it in a 100ml measuring bottle, add an appropriate amount of water for injection to dissolve, then add 1.0g of GSK2239633, add an appropriate amount of sodium hydroxide (1.5ml, 0.5mol/L), shake To dissolve, add 10.0 g of polyoxyethylene lauryl ether, add water for injection close to the mark, shake well, adjust the pH to 8.0 with sodium bicarbonate, add 0.3 g of sodium chloride, add 2.5 g of β-cyclodextrin, add Water for injection was adjusted to the mark, shaken, and the liquid was filtered with a 0.22 μm microporous membrane to obtain a nasal spray liquid of a CCR4 inhibitor, with a pH of about 8.5. It is then filled into single-dose and multi-dose nasal spray devices, respectively, and packaged for later use.

Example 6: Preparation of CCR4 inhibitor nasal spray 6

Weigh 0.03g of sodium methylparaben, put it in a 100ml measuring bottle, add an appropriate amount of water for injection to dissolve, then add 1.0g of GSK2239633, add an appropriate amount of sodium hydroxide (1.5ml, 0.5mol/L), shake To dissolve, add 15.0g of propylene glycol, add water for injection close to the mark, shake well, add 0.08g of citric acid and 0.8g of sodium citrate, adjust the pH to 8.0, add 1.5g of β-cyclodextrin, and add water for injection to the mark , shake well, filter the medicinal solution with a 0.22 μm microporous membrane, and obtain a nasal spray medicinal solution of a CCR4 inhibitor with a pH of about 8.5. It is then filled into single-dose and multi-dose nasal spray devices, respectively, and packaged for later use.

Example 7: Preparation of CCR4 inhibitor nasal spray 7

Weigh 0.03g of sodium methylparaben, put it in a 100ml measuring bottle, add an appropriate amount of water for injection to dissolve, then add 250mg of GSK2239633, add an appropriate amount of sodium hydroxide (1.5ml, 0.5mol/L), shake to make Dissolve, add water for injection close to the mark, shake well, adjust the pH to 8.0 with sodium dihydrogen phosphate, add 0.3 g of sodium chloride, add water for injection to the mark, shake well, filter the medicinal solution with a 0.22 μm microporous membrane, A nasal spray solution of a CCR4 inhibitor was obtained with a pH of about 8.5. It is then filled into single-dose and multi-dose nasal spray devices, respectively, and packaged for later use.

Example 8: Preparation of CCR4 inhibitor nasal spray 8

Weigh 0.03g of sodium methylparaben, put it in a 100ml measuring bottle, add an appropriate amount of water for injection to dissolve, then add 250mg of GSK2239633, add an appropriate amount of sodium hydroxide (1.5ml, 0.5mol/L), shake to make Dissolve, add 15.0g of propylene glycol, add water for injection to the mark, shake well, adjust the pH value to 8.5 with sodium bicarbonate, add 0.3g of sodium chloride, add water for injection to the mark, shake well, use a 0.22μm microporous filter membrane Filter the medicinal solution to obtain a nasal spray medicinal solution of CCR4 inhibitor, the pH is about 9.0. It is then filled into single-dose and multi-dose nasal spray devices, respectively, and packaged for later use.

Example 9: Preparation of CCR4 inhibitor nasal spray 9

Weigh 0.03g of sodium methylparaben, put it in a 100ml measuring bottle, add an appropriate amount of water for injection to dissolve, then add 1.0g of GSK2239633, add an appropriate amount of sodium hydroxide (1.5ml, 0.5mol/L), shake To dissolve, add 15.0g of propylene glycol, add water for injection close to the mark, shake well, add citric acid and sodium citrate (1:10) buffer, adjust the pH to 8.5, add 0.3g of sodium chloride, add β-cyclodextrin 1.5 g of sperm, add water for injection to the mark, shake well, filter the liquid with a 0.22 μm microporous membrane, and obtain a nasal spray liquid of CCR4 inhibitor, with a pH of about 9.0. It is then filled into single-dose and multi-dose nasal spray devices, respectively, and packaged for later use.

Experimental Example 10: In vitro ciliary toxicity test

This experiment evaluated the nasal ciliary toxicity of the previously prepared formulations with different concentrations of CCR4 inhibitor. In the experiment, sodium deoxycholate, which is recognized in the literature as having severe ciliary toxicity, was used as a positive control, and normal saline was used as a blank control.

1. Experimental samples

CCR4 inhibitor nasal spray 1-9 prepared in Example 1-9;

Positive control (1% sodium deoxycholate solution);

Blank control (normal saline).

2. Experimental method

Lasting time of eilliary movement (LTCM) was measured by in vitro method of cilia on toad palate.

Fix the toad, cut the upper palate mucosa with surgical scissors, put it in normal saline, wash away blood clots and sundries, and quickly use ophthalmic scissors to cut about 3x3mm ² of the upper palate mucosa, and lay the mucociliary side up on a glass slide , add 0.1mL liquid medicine on the mucosal surface, cover with a cover glass gently, observe the movement of cilia with a microscope until the movement of cilia stops, and record the time from drug administration to stop of movement of cilia.

Carefully wash the liquid on the mucous membrane with normal saline, continue to observe whether the movement of the cilia is restored, and record the continuous movement time of the recovery sample.

3. Experimental results

as in Table 1 below.

Table 1 Evaluation results of ciliary toxicity

Sample information	LTCM (minutes)	Relative percentage (%)	Recovery time (minutes)
normal saline	648±36	-	-
positive control	7±2	1.1	-
Example 1	427±13	65.9	126±9
Example 2	456±20	70.4	128±9
Example 3	487±21	75.2	131±8
Example 4	356±17	54.9	186±9
Example 5	347±16	53.5	178±11
Example 6	367±17	56.6	185±12
Example 7	433±12	66.8	121±8
Example 8	455±17	70.2	118±4
Example 9	360±12	55.6	167±8

4. Experimental conclusion

1) The mucosal surface of the blank control group was intact under the microscope, and the cilia were active.

2) In the positive control group, the cilia on the mucosal surface fell off under the microscope, and the cilia did not swing and could not recover.

3) The CCR4 inhibitor nasal sprays prepared in Examples 1-9 have certain ciliary toxicity, but they can all recover after a certain period of time, and the toxicity is reversible.

4) The ciliary toxicity of GSK2239633 increased with increasing concentration, but was also reversible.

5) The ciliary toxicity of other components in the preparation is relatively small, and basically has no obvious effect.

Example 11 In vivo ciliary toxicity evaluation

1. Experimental samples

CCR4 inhibitor nasal sprays 1-9 prepared in Examples 1-9.

The blank control was normal saline.

2. Experimental method

The toad was fixed in the supine position, the mouth was opened, and 0.5 ml of the drug solution was dripped on the upper palate mucosa to completely immerse the upper palate. After contacting for 30 minutes, the drug was washed with normal saline, and the upper palate mucosa was separated for microscope observation of ciliary movement. The mucosa was washed immediately after separation, flattened on a glass slide, dripped with normal saline on the surface of the mucosa, covered with a cover glass, and observed and recorded the duration of ciliary movement as described above.

3. Experimental results

as in Table 2 below.

Table 2 In vivo ciliary toxicity evaluation results

Sample information	LTCM (minutes)	Relative percentage (%)
normal saline	816±37	-
Example 1	760±23	93.1
Example 2	756±20	92.6
Example 3	787±25	96.4
Example 4	666±17	81.6
Example 5	677±18	83.0
Example 6	691±17	84.7
Example 7	753±22	92.3
Example 8	775±19	95.0
Example 9	660±13	80.9

The ciliary toxicity of the CCR4 inhibitor nasal spray prepared in Examples 1-9 was evaluated by the in vivo method. It was found that the ciliary toxicity of each preparation was lower than that in the in vitro experiment, and the continuous movement time of the cilia was longer than that in the in vitro method. The preparations of the above examples all have good ciliary safety and are suitable for nasal administration.

Experimental Example 12: Evaluation of Bioavailability

1. Experimental samples

CCR4 inhibitor nasal spray, prepared in Examples 1-9.

Preparation of CCR4 inhibitor injection: in a 100ml volumetric bottle, add an appropriate amount of water for injection, then add 250mg of GSK2239633, add an appropriate amount of sodium hydroxide (1.5ml, 0.5mol/L), shake to dissolve, shake well, and adjust with sodium bicarbonate The pH value was adjusted to 8.5, water for injection was added to the scale, shaken well, and the liquid was filtered with a 0.22 μm microporous membrane to prepare a CCR4 inhibitor injection liquid.

Preparation of oral preparation of CCR4 inhibitor: in a 100ml measuring bottle, add an appropriate amount of water for injection, then add GSK2239633 250mg, add an appropriate amount of sodium hydroxide (1.5ml, 0.5mol/L), shake to dissolve, shake well, and use sodium bicarbonate Adjust the pH value to 10, add water for injection to the scale, shake well, and filter the medicinal solution with a 0.22 μm microporous membrane to prepare an oral medicinal solution of CCR4 inhibitor.

2. Experimental method

Route of administration: nasal spray, oral, injection.

Dosage: oral 5mg/kg, nasal spray, injection 1mg/kg.

Dosing regimen:

The experiment adopted a three-period, self-controlled crossover design. 27 male beagle dogs were randomly divided into 9 groups. After a one-week washout period in each period, CCR4 inhibitors were administered orally, by injection, and by nasal spray, respectively. relative bioavailability. At 0min before administration and 0.033, 0.083, 0.167, 0.5, 1, 2, 3, 4, 6, and 8 hours after administration, about 1ml of blood was alternately collected from the veins of the limbs, and transferred to a 1.5ml EP tube at 3000 rpm/ minutes, centrifuged for 15 minutes, and separated about 100 μL of plasma.

Sample processing:

Take 100 μL of beagle dog plasma, add 100 μL dexmedetomidine (internal standard,

50ng/ml), add 0.5ml methanol, after sufficient shaking, centrifuge at 12,000 rpm for 6 minutes, transfer 200μl of supernatant to a sample bottle, and take 10μl for LC/MS/MS analysis and determination.

Table 3: Relative bioavailability studies of CCR4 inhibitors

Sample information	bioavailability(%)
Injectable preparations	-
Oral preparation	25.0±0.2
Example 1	90.1±0.4
Example 2	89.3±0.5
Example 3	88.7±0.3
Example 4	91.2±0.5
Example 5	90.3±0.4
Example 6	88.6±0.4
Example 7	85.4±0.3
Example 8	90.2±0.5
Example 9	92.2±0.4

3. Experimental results

The results showed that after three routes of nasal spray, oral and intravenous administration of CCR4 inhibitor, with injection as control, the relative bioavailability of the oral preparation was 25.0%, the relative bioavailability of the nasal spray preparation was 85.4-92.2%, The bioavailability is increased by 3.47-3.69 times, effectively overcoming the deficiency of CCR4 inhibitors.

The present invention overcomes the defects of poor solubility and bioavailability of the CCR4 inhibitor GSK2239633, and is the first in the field to prepare and provide a CCR4 inhibitor preparation that is more suitable for clinical applications. Specifically, the present invention provides a technique for improving the bioavailability of GSK2239633 through mucosal administration routes such as nasal mucosal administration and a usable pharmaceutical preparation, which solves the problem that the CCR4 inhibitor in the prior art has low solubility and affects the bioavailability of active pharmaceutical ingredients. Release, resulting in too low bioavailability, there is a problem that the expected effective plasma concentration cannot be achieved in the body at the maximum safe dose. The CCR4 inhibitor pharmaceutical preparation provided by the present invention also has the advantages of no local irritation and safe use.

The above is the description of the present invention, and should not be construed as limiting the present invention. Unless otherwise indicated, the practice of this invention will employ conventional techniques of organic chemistry, polymer chemistry, biotechnology, etc., it being apparent that the invention may be practiced otherwise than as specifically described in the foregoing specification and examples. Other aspects and modifications within the scope of the invention will be apparent to those skilled in the art to which this invention pertains. Many modifications and variations are possible in light of the teachings of the present invention and are therefore within the scope of the present invention.

Claims (20)

1. A dosage form for mucosal administration comprising a compound of formula I or a pharmaceutically acceptable salt, solvate or prodrug thereof:

   
2. The mucosal administration dosage form of claim 1, which is a nasal administration dosage form.
3. 2. The dosage form for mucosal administration according to claim 1 or 2, which is a solution composition, eg a solution composition suitable for spraying.
4. A dosage form for mucosal administration according to claim 3, comprising a cosolvent such as sodium hydroxide, sodium methoxide, potassium hydroxide, sodium bicarbonate, sodium dihydrogen phosphate, sodium citrate, α-, β- or γ-ring Any one or more of the dextrins, preferably, the co-solvent contained therein is sodium hydroxide and/or β-cyclodextrin, and more preferably, the co-solvent contained therein is sodium hydroxide.
5. The mucosal administration dosage form of claim 4, which comprises the cosolvent in an amount of about 0.25-1.5 mol/L, preferably 0.5-1.0 mol/L.
6. A dosage form for mucosal administration according to claim 3, comprising a pH adjusting agent such as phosphoric acid, hydrochloric acid, citric acid, tartaric acid, acetic acid, maleic acid, sodium hydroxide, sodium carbonate, sodium bicarbonate, sodium citrate, phosphate buffer Any one or more of the solution, acetate buffer and citrate buffer, preferably, the pH adjuster it contains is phosphate and/or citrate buffer.
7. A dosage form for mucosal administration according to claim 4, comprising a preservative such as methylparaben, ethylparaben, propylparaben, butylparaben, methylparaben Sodium, Sodium Ethylparaben, Sodium Propylparaben, Sodium Butylparaben, Sorbic Acid, Sodium Sorbate, Benzoic Acid, Sodium Benzoate, Benzyl Alcohol, Benzalkonium Bromide, Benzalkonium Chloride Any one or more of ammonium, trichlorobutanol, resorcinol and sodium EDTA, preferably, the preservative is sodium methyl parahydroxybenzoate.
8. 4. The dosage form for mucosal administration according to claim 4, which comprises an osmotic pressure regulator, such as any one or more of sodium chloride, potassium nitrate, boric acid and glucose, preferably sodium chloride.
9. The mucosal administration dosage form of claim 4, which comprises a penetration enhancer such as selected from (1) alcohols, esters, dimethyl sulfoxide and homologues in organic solvents; (2) fatty acids, fatty alcohols and cholate salts (3) surfactants such as cationic, anionic, nonionic and lecithin; (4) terpenes; and (5) any one or more of keratin moisturizing and softening agents, preferably propylene glycol and or polyoxyethylene lauryl ether.
10. 9. The mucosal administration dosage form of any one of claims 1-9, which has a pH of about 6.5-9.5; preferably about 7.0-8.5.
11. The mucosal administration dosage form of any one of claims 1-10, which is present in an amount of about 1.25 mg to 10.0 mg of the compound of formula I or a pharmaceutically acceptable salt, solvate or prodrug thereof per 100 ml of the dosage form g, preferably about 2.5 mg to 5 g, and preferably about 5 mg to 1 g.
12. The mucosal administration dosage form of any one of claims 1-11, which is a dosage form for single-dose or multiple-dose administration, wherein the compound of formula I, or a pharmaceutically acceptable salt, solvate thereof, is administered at each dose Or the amount of prodrug is about 2.5 μg to 10.0 mg, preferably about 5.0 μg to 1.0 mg.
13. 12. The mucosal administration dosage form of any one of claims 1-12, which is a solution for spraying with a spray particle size of about 5-200 [mu]m; preferably about 10-150 [mu]m.
14. 14. The mucosal administration dosage form of any one of claims 1-13 for use in the treatment of a CCR4 mediated disease.
15. The mucosal administration dosage form of claim 14, wherein the CCR4-mediated disease is selected from the group consisting of: (1) respiratory diseases, such as airway obstructive diseases, including asthma, including bronchial asthma, allergic asthma, endogenous asthma, Extrinsic asthma, exercise-induced asthma, drug-induced (including aspirin and NSAID-induced) and dust-induced asthma, intermittent and persistent asthma; chronic obstructive pulmonary disease (COPD); bronchitis, including infectious Bronchitis and eosinophilic bronchitis; emphysema; bronchiectasis; cystic fibrosis; hypersensitivity pneumonitis; pulmonary fibrosis, including cryptogenic fibrotic alveolitis, idiopathic interstitial pneumonia, antineoplastic therapy and Fibrosis complicated by chronic infection; vasculitis and thrombotic disease of the pulmonary vessels and pulmonary hypertension; rhinitis: acute rhinitis and chronic rhinitis, including drug-induced rhinitis and vasomotor rhinitis; perennial allergic rhinitis and seasonal allergic rhinitis Rhinitis, including neurorhinitis; acute viral infections, including the common cold and infections caused by respiratory syncytial virus, influenza, coronavirus, and adenovirus,

    Preferably, wherein the CCR4 mediated diseases are asthma, COPD and rhinitis.
16. A method of preparing a dosage form for mucosal administration as previously described comprising a compound of formula I or a pharmaceutically acceptable salt, solvate or prodrug thereof:

    

    .
17. The method of claim 16, wherein the dosage form for mucosal administration is a nasal dosage form, such as a solution composition, wherein the method comprises compounding a compound of formula I or a pharmaceutically acceptable salt, solvate or prodrug thereof Steps for mixing with co-solvents.
18. The method of claim 17, wherein the concentration of the cosolvent added is about 0.1-3.0 mol/L, preferably about 0.2-2.0 mol/L, more preferably about 0.3-1.0 mol/L, for example about 0.5 mol/L , and about 0.5-3 parts by volume, preferably about 1-2 parts by volume, for example about 1.5 parts by volume of cosolvent per 100 parts by volume of the mucosal administration dosage form.
19. 18. The method of claim 17, comprising the step of adjusting the pH of the mucosal administration dosage form to about 6.5-9.5; preferably about 7.0-8.5.
20. 18. The method of claim 17, further comprising the step of adding one or more of preservatives, osmotic pressure regulators, penetration enhancers.