Clathrate compound containing baricitinib as well as preparation method and application of clathrate compound

Abstract

The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a baricitinib-containing clathrate compound as well as a preparation method and application thereof. The invention provides a baricitinib clathrate compound. The baricitinib clathrate compound comprises cyclodextrin and an active ingredient clathrated in the cyclodextrin, the baricitinib is included by cyclodextrin, so that the solubility and stability of the baricitinib are remarkably improved, and meanwhile, the preparation process is simplified. The local administration preparation of the baricitinib clathrate compound can quickly reach the focus, enhances the local administration efficiency, makes the pharmacological activity fully exerted, reduces the adverse reaction of the whole body, effectively reduces the level of inflammatory factors, and provides a safe and efficient new treatment method for local lesions such as rhinitis and nasosinusitis.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical preparations, and in particular relates to a baricitinib-containing inclusion compound and a preparation method and application thereof. Background Art

[0002] In the pharmaceutical field, baricitinib, as an effective small molecule drug, has been widely used to treat a variety of inflammatory diseases. However, baricitinib has significant deficiencies in water solubility, and its solubility in water is extremely low (0.02 mg / ml), which limits its application in topical preparations, especially in the treatment of local lesions such as rhinitis. Baricitinib in the form of oral tablets is difficult to reach the lesions quickly and effectively, affecting the therapeutic effect.

[0003] In addition, systemic administration may lead to uneven distribution of drugs in the body, increase the risk of adverse reactions, and reduce patients' treatment compliance. Therefore, improving the water solubility of baricitinib and developing effective local administration preparations have become technical issues that need to be solved urgently.

[0004] In the prior art, although there are some methods to try to improve the solubility of poorly soluble drugs, such as using surfactants, solid dispersion technology, etc., these methods often have problems such as complex preparation, high cost or insufficient stability. At the same time, these technologies may introduce new side effects or reduce the bioavailability of drugs while improving solubility.

[0005] In order to overcome the limitations of the prior art, the present invention proposes a new technical solution, namely, using cyclodextrin to include baricitinib, in order to solve the problems of low solubility and low efficiency of local administration, thereby improving the clinical application value of baricitinib. Summary of the invention

[0006] The object of the present invention is to provide a baricitinib inclusion compound. The baricitinib inclusion compound provided by the present invention has high solubility, thereby improving the efficiency of local administration.

[0007] In order to achieve the above object, the present invention provides the following technical solutions:

[0008] The present invention provides a baricitinib inclusion compound, which comprises cyclodextrin and a baricitinib active ingredient included in the cyclodextrin.

[0009] A method for preparing a baricitinib inclusion complex comprises: mixing a baricitinib active ingredient, cyclodextrin and water, and performing an inclusion reaction to obtain the baricitinib inclusion complex.

[0010] A baricitinib inclusion compound solution comprises the baricitinib inclusion compound and water, wherein the content of the baricitinib inclusion compound is 0.01% to 0.5% based on the mass of baricitinib.

[0011] Preferably, the baricitinib inclusion complex solution further comprises a preservative, a pH regulator, a transdermal enhancer or a viscosity regulator.

[0012] The present invention also provides the use of the above-mentioned baricitinib inclusion complex or baricitinib inclusion complex solution, which is used in the preparation of drugs for preventing and treating allergic rhinitis, eosinophilic sinusitis, eosinophilic chronic sinusitis with nasal polyps, non-eosinophilic chronic sinusitis with nasal polyps, chronic sinusitis without nasal polyps or chronic sinusitis with nasal polyps.

[0013] Preferably, the use is for preparing a drug for reducing the levels of IL13, IFNγ and IL17A in polyp tissues of CRSwNP patients.

[0014] Preferably, the use is for preparing a drug for allergic symptoms of allergic rhinitis

[0015] Preferably, the medicament is a topical preparation.

[0016] Preferably, the use is to locally inhibit the expression levels of IL-4 and IL-13 in the nasal mucosa.

[0017] The present invention also provides the use of the above-mentioned baricitinib inclusion solution in the preparation of a drug for treating rhinitis, sinusitis, asthma, rheumatoid arthritis, ankylosing spondylitis, psoriasis, atopic dermatitis, systemic lupus erythematosus, ulcerative enteritis or alopecia areata.

[0018] Preferably, the dosage form of the drug includes spray, nasal drops, inhalation, oral solution, injection or gel.

[0019] Beneficial effects:

[0020] (1) The present invention significantly improves the solubility of baricitinib through cyclodextrin inclusion, and the solubility in water is increased from 0.02 mg / ml to 5 mg / ml. This allows the local administration preparation containing the baricitinib inclusion complex to quickly reach the lesion, so that its pharmacological activity can be fully exerted; and it can reduce the adverse reactions of the drug during systemic administration and improve patient compliance.

[0021] (2) The present invention simplifies the inclusion process by directly adding baricitinib in powder form into a cyclodextrin solution and performing a micronization process, thereby avoiding the use of organic solvents and removal steps.

[0022] (3) The baricitinib described in the present invention has good stability. When the aqueous solution of the baricitinib inclusion complex is placed in the dark at 40° C. for 6 months, the properties and content of the solution do not change significantly, indicating good stability.

[0023] (4) The baricitinib inclusion complex solution of the present invention can significantly reduce the levels of inflammatory factors, such as IL13, IFNγ and IL17A, in the polyp tissue of CRSwNP patients, thereby alleviating the inflammatory response.

[0024] (5) The baricitinib inclusion complex of the present invention has a therapeutic effect on eosinophilic sinusitis by reducing the level of inflammatory factors, and the inclusion complex does not reduce the efficacy of the drug.

[0025] (6) The baricitinib inclusion compound of the present invention can reduce the allergic reaction of allergic rhinitis. Moreover, the baricitinib inclusion compound solution is only effective locally when administered nasally, has little effect on the whole body, and has high safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0027] Figure 1 Immunofluorescence was used to detect the expression of p-JAK1 and p-JAK2 in tissues of patients with sinusitis and sinusitis with nasal polyps, including EOS-CRSwNP: patients with eosinophilic chronic sinusitis with nasal polyps, NonEOS-CRSwNP: patients with non-eosinophilic chronic sinusitis with nasal polyps, CRSsNP: patients with chronic sinusitis without nasal polyps, and NM: normal controls.

[0028] Figure 2 The effect of baricitinib on the expression of IL-13, IFN-γ and IL17A in polyp tissues of patients with sinusitis and nasal polyps (Mean±SD, n=4, ns=not significant, *P<0.05).

[0029] Figure 3 Real-time PCR was used to detect the mRNA levels of inflammatory factors IL4 and IL13 in the mouse nasal mucosa.

[0030] Figure 4 This is the effect of baricitinib on the frequency of nose scratching and sneezing in AR mice.

[0031] Figure 5 Real-time PCR was used to detect the mRNA levels of inflammatory factors in the mouse nasal mucosa.

[0032] Figure 6 Real-time PCR was used to detect the mRNA levels of inflammatory factors in mouse spleen. DETAILED DESCRIPTION

[0033] The present invention provides a baricitinib inclusion compound and a preparation method thereof, and a baricitinib inclusion compound solution and a preparation method thereof, and also provides specific applications of the baricitinib inclusion compound and the solution.

[0034] The meanings of the abbreviations in the present invention are:

[0035] AR: allergic rhinitis;

[0036] EOS-CRS: eosinophilic sinusitis;

[0037] EOS-CRSwNP: patients with eosinophilic chronic sinusitis with nasal polyps;

[0038] NonEOS-CRSwNP: patients with noneosinophilic chronic sinusitis with nasal polyps;

[0039] CRSwNP: chronic rhinosinusitis with nasal polyps patients

[0040] CRSsNP: chronic rhinosinusitis without nasal polyps patients;

[0041] NM: normal control;

[0042] JAK: selective tyrosine protein kinase;

[0043] JAK-STAT: a series of chain reactions between intracellular proteins.

[0044] P-JAK1: phosphorylation tyrosine protein kinase-1

[0045] P-JAK2: phosphorylation tyrosine protein kinase-2

[0046] In order to further illustrate the present invention, the technical solution provided by the present invention is described in detail below in conjunction with the accompanying drawings and embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0047] The production processes, experimental methods or detection methods involved in the embodiments of the present invention, unless otherwise specified, are all conventional methods in the prior art, and their names and / or abbreviations are all conventional names in the field, and are very clear and unambiguous in the relevant application fields. Technical personnel in the field can understand the conventional process steps based on the names and apply the corresponding equipment, and implement them according to conventional conditions or the conditions recommended by the manufacturer.

[0048] The various instruments, equipment, raw materials or reagents used in the embodiments of the present invention are not particularly limited in terms of their sources, and are all conventional products that can be purchased through regular commercial channels, or can be prepared according to conventional methods well known to those skilled in the art.

[0049] Example 1

[0050] Preparation of Baricitinib inclusion complex

[0051] Mix 50 g of sodium sulfobutyl β-cyclodextrin and about 70 ml of pure water to obtain a first solution;

[0052] Mix 500 mg of baricitinib and 10 mL of pure water to obtain a second suspension; slowly drop the obtained second mixed solution into the first mixed solution under stirring, and carry out inclusion reaction at 40°C for 2 hours; filter the obtained inclusion solution, collect the filtrate, the baricitinib content is 5 mg / ml, dry the filtrate under vacuum conditions to obtain 50.12 g of baricitinib inclusion complex, and then grind it mechanically to obtain it.

[0053] Example 2

[0054] Preparation of Baricitinib inclusion complex solution

[0055] 10.09 g (containing 100 mg of baricitinib) of the inclusion compound prepared according to Example 1 was mixed with pure water, stirred at 25°C to fully dissolve it, then 0.2 g of potassium sorbate was added, and the pH value of the mixture was adjusted to 4.50 with dilute hydrochloric acid, and the volume was made up to 50 ml with pure water to obtain a baricitinib inclusion compound solution with a concentration of 2 mg / mL. After filtering through a 0.45 μm filter membrane, it was sealed and stored in a light-proof manner.

[0056] The obtained baricitinib inclusion complex solution is filled into corresponding packaging containers to prepare nasal sprays or nasal drops.

[0057] The prepared product was subjected to performance test

[0058] Test Example 1

[0059] Stability test of baricitinib inclusion complex aqueous solution at 40°C and solubility test at 25°C:

[0060] The 2 mg / mL aqueous solution of baricitinib inclusion complex obtained in Example 2 was dispensed into glass ampoules and placed away from light at 40° C. The properties of the solution were observed at different times, and the related substances and their contents were detected. The test results are shown in Table 1. The related substances include: potassium sorbate and baricitinib inclusion complex.

[0061] Table 1 Storage stability of aqueous solutions containing baricitinib inclusion complexes

[0062] Storage conditions Solution properties Related substances (%) content(%) 40℃, October Clarity and transparency 0.46 100.00 40℃, January Clarity and transparency 0.45 99.76％ 40℃, February Clarity and transparency 0.60 99.50％ 40℃, March Clarity and transparency 0.71 99.31％ 40℃, June Clarity and transparency 0.73 99.07％

[0063] The results showed that when the aqueous solution of baricitinib inclusion complex was placed in the dark at 40°C for 6 months, there was no significant change in the properties and content of the solution, and the related substances increased slightly, indicating that the sample had good stability under this condition.

[0064] At 25° C., the baricitinib inclusion complex obtained in Example 1 was dissolved in water in excess according to the method of Example 2 to prepare a saturated aqueous solution. The concentration of the saturated aqueous solution was measured to be 5 mg / ml.

[0065] Test Example 2

[0066] Expression of JAK-STAT in tissues of patients with sinusitis and sinusitis with nasal polyps Nasal polyp tissues of patients with clinical eosinophilic and non-eosinophilic chronic sinusitis with nasal polyps, ethmoid sinus mucosa tissues of patients with chronic sinusitis, and lower nail tissues of normal subjects were obtained. The results showed that P-JAK1 was highly expressed in mucosal tissues of patients with various types of rhinitis, especially in the mucosa of nasal polyps of NonEOS-CRSwNP patients, where the expression was significantly increased. P-JAK2 was also highly expressed in mucosal tissues of patients with various types of rhinitis, and the increase was most significant in the mucosa of nasal polyps of EOS-CRSwNP patients. It was also highly expressed in inflammatory cells in the mucosal tissues of EOS-CRSwNP and CRSsNP patients, such as Figure 1 shown.

[0067] Test Example 3

[0068] Baricitinib inclusion complex solution inhibits the expression of IL13, IFNγ and IL17A in nasal polyp tissues of patients with CRSwNP

[0069] Human nasal polyp tissue was cultured using the air-liquid interface method. Different concentrations of baricitinib inclusion complex solutions were added to each well of the Transwell chamber to make the final concentrations of 1 mg / L, 10 mg / L, and 100 mg / L, respectively. After 6 hours of action, the expression levels of IFNγ, IL13, and IL17A in the tissue were detected by real-time PCR.

[0070] like Figure 2As shown in the results, the baricitinib inclusion complex solution can significantly reduce the levels of IL13, IFNγ and IL17A in the polyp tissues of CRSwNP patients in a dose-dependent manner. IFN-γ, as a proinflammatory cytokine, participates in the early response and amplification of the inflammatory response, and IL-13 can promote the production of IFN-γ. IL-13 is a key cytokine that induces and maintains type 2 responses and is associated with a variety of atopic diseases, such as atopic dermatitis (AD), asthma, and chronic sinusitis with nasal polyposis. Several therapeutic agents that selectively target type 2 cytokines, such as Dupilumab, have shown efficacy in one or more of these atopic diseases. As a proinflammatory factor, IL17A can induce proinflammatory cells to secrete chemokines or cytokines after binding to the receptor, and participate in the acute phase of inflammation. The above results indicate that baricitinib can reduce the expression of inflammatory factors in the lesion tissues of CRSwNP patients, thereby alleviating the inflammatory response.

[0071] Test Example 4

[0072] To investigate the effects of baricitinib and baricitinib inclusion complex solution on eosinophilic sinusitis (EOS-CRS)

[0073] Female C57BL / 6 wild-type mice (4-6 weeks old) were intranasally administered 1 mg / ml papain solution (Sigma-Aldrich) once a day on days 0-2 and 7-10, with 10 μl per nostril each time to induce a sinusitis model.

[0074] Fifty-six mice were randomly divided into seven groups, with eight mice in each group. Except for the control group, the mice in each group were given papain nasal drops to establish the sinusitis model. The mice in the control group were given the same volume of PBS solution. Each treatment group was given different concentrations of baricitinib suspension and baricitinib inclusion solution by nasal drops on days 4-10, once a day, 10 μl per side each time, a total of 20 μl. On day 11, the real-time PCR method was used to detect the expression levels of IL4 and IL13 genes in the nasal mucosa of mice.

[0075] Type II inflammatory cytokines such as IL4 and IL13 play an important role in the pathogenesis of chronic sinusitis. Figure 3 The results showed that compared with the blank control group, the inflammatory factors IL4 and IL13 in the nasal mucosa of the ECRS group mice were significantly increased after papain nasal drops, indicating that this modeling method is ideal. The baricitinib suspension and baricitinib inclusion solution in each administration group can inhibit the expression of IL4 and IL13 genes to varying degrees, and at 2 mg / ml, the inhibitory effect of the baricitinib inclusion solution on IL4 and IL13 is equivalent to that of the uninclusion baricitinib, and the inhibitory effect on IL13 is dose-dependent. The above results suggest that baricitinib can play a therapeutic role in eosinophilic sinusitis by reducing the level of inflammatory factors. Figure 3The control in the table indicates blank control, ECRS indicates eosinophilic sinusitis, BAR1, 2, 5 indicate 1 mg / ml, 2 mg / ml, 5 mg / ml baricitinib inclusion complex solutions, BAR2q indicates 2 mg / ml baricitinib suspension, and TAC indicates triamcinolone acetonide as a positive control.

[0076] Test Example 5

[0077] To investigate the effect of baricitinib inclusion complex solution on allergic rhinitis (AR)

[0078] Female C57BL / 6 wild-type mice (4-6 weeks old) were intraperitoneally injected (ip) with 200 μl of sensitizer (0.5 μl of 50 μg / μl ovalbumin solution + 100 μl PBS + 100 μl alum adjuvant) on days 1 and 7, respectively. On days 14-21, the mice were challenged with 50 μg / μl ovalbumin solution intranasally (in) once a day, 10 μl per nostril each time. Samples were collected on day 23 to induce the establishment of an AR model.

[0079] 56 mice were randomly divided into 7 groups, 8 mice in each group. Except for the control group, the other mice were sensitized and stimulated with ovalbumin solution to induce AR formation, and the control group mice were given the same volume of PBS solution. The treatment group was given 1mg / ml, 2mg / ml, and 5mg / ml baricitinib inclusion complex solution by intranasal drops on days 11-21, and another group was set up to investigate the blank excipient solution, once a day, 10μl per nostril each time, and triamcinolone acetonide was used as a positive control.

[0080] After the last ovalbumin solution challenge, the number of nose scratching and sneezing in each group of mice within 5 minutes was recorded. On the 23rd day, the mice were killed by CO2 anesthesia, and the nasal septum mucosa, sinus mucosa and spleen were obtained to detect the expression levels of IL4 and IL13.

[0081] Experimental results:

[0082] Effects of baricitinib inclusion complex solution on AR allergic symptoms

[0083] like Figure 4 As shown, the number of nose scratching and sneezing in AR mice was significantly increased compared with normal mice. After 10 days of nasal drops treatment with baricitinib inclusion complex solution, 2mg / ml and 5mg / ml baricitinib inclusion complex solutions could significantly reduce the number of nose scratching and sneezing in the model group mice and relieve allergic symptoms, and the blank excipient solution had no significant effect on the mice. Figure 4 The control represents blank control, PBS is buffer solution, and TAC represents triamcinolone acetonide as positive control.

[0084] Effects of baricitinib inclusion complex solution on the expression levels of IL-4 and IL-13 in the nasal mucosa and spleen of AR mice

[0085] like Figure 5 As shown, 2 mg / ml of baricitinib inclusion complex solution can inhibit the expression of inflammatory factor IL4 in the nasal mucosa of AR mice, and 5 mg / ml has a significant inhibitory effect on both IL4 and IL13, indicating that baricitinib shows a significant inhibitory effect on type II inflammation in allergic rhinitis in mice, and this inhibitory effect is comparable to that of the classic intranasal hormone drug triamcinolone acetonide.

[0086] Intranasal treatment with baricitinib solution and triamcinolone acetonide solution had no significant effect on spleen inflammatory factors in each group of mice ( Figure 6 ), indicating that the baricitinib inclusion solution is only effective locally when administered intranasally, has little effect on the whole body, and is relatively safe. Figure 5 and 6 The control represents blank control, PBS is buffer solution, and TAC represents triamcinolone acetonide as positive control.

[0087] It can be seen from the above embodiments that the present invention significantly improves the solubility and stability of baricitinib by cyclodextrin inclusion, simplifies the preparation process, enhances the efficiency of local administration, reduces systemic adverse reactions, and effectively reduces the level of inflammatory factors, thereby providing a safe and efficient new treatment method for local lesions such as rhinitis and sinusitis.

[0088] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A baricitinib inclusion compound, characterized in that: The inclusion compound comprises cyclodextrin and the baricitinib active ingredient included in the cyclodextrin.

2. A method for preparing a baricitinib inclusion compound, characterized in that: The method comprises: mixing the active ingredient of baricitinib, cyclodextrin and water, and performing an inclusion reaction to obtain the baricitinib inclusion compound.

3. A baricitinib inclusion compound solution, characterized in that The solution comprises a baricitinib inclusion complex and water, wherein the content of the baricitinib inclusion complex is 0.01% to 0.5% based on the mass of baricitinib; the baricitinib inclusion complex is the inclusion complex according to claim 1 or the inclusion complex prepared by the preparation method according to claim 2.

4. The solution according to claim 3, characterized in that The baricitinib inclusion compound solution further comprises a preservative, a pH regulator, a transdermal enhancer or a viscosity regulator.

5. Use of the inclusion compound according to claim 1, the inclusion compound prepared by the preparation method according to claim 2, or the baricitinib inclusion compound solution according to any one of claims 3 to 4, characterized in that: The use is for preparing medicines for preventing and treating allergic rhinitis, eosinophilic sinusitis, eosinophilic chronic sinusitis with nasal polyps, non-eosinophilic chronic sinusitis with nasal polyps, chronic sinusitis without nasal polyps or chronic sinusitis with nasal polyps.

6. The use according to claim 5, characterized in that The use is for preparing a drug for reducing the levels of IL13, IFNγ and IL17A in polyp tissues of CRSwNP patients.

7. The use according to claim 5, characterized in that The use is for preparing a medicine for allergic symptoms of allergic rhinitis.

8. The use according to claim 5, characterized in that The drug is a preparation for topical administration.

9. The use according to claim 8, characterized in that The use is to locally inhibit the expression levels of IL-4 and IL-13 in the nasal mucosa.

10. Use of the baricitinib inclusion compound solution according to any one of claims 3 to 4 in the preparation of a drug for treating rhinitis, sinusitis, asthma, rheumatoid arthritis, ankylosing spondylitis, psoriasis, atopic dermatitis, systemic lupus erythematosus, ulcerative enteritis or alopecia areata.

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