KR20190043865A - Depot formulation for injection comprising entecavir fatty acid esters analogs - Google Patents

Abstract

The present invention relates to an injectable depot formulation comprising fatty acid ester derivatives of entecavir. According to the present invention, entecavir, which is orally administered conventionally, is made into a prodrug and prepared as a depot formulation to be administered through intramuscular or subcutaneous injection. To design an injectable formulation capable of maintaining a plasma drug concentration of entecavir at 3-week time point or even later after administering fatty acid ester derivatives of entecavir, it was essential to use fine particles having sizes within the range of 1-50 μm. Furthermore, it was found that, as particle sizes of suspended particles of fatty acid ester derivatives of entecavir decrease, the duration for which the plasma drug concentration is maintained decreases as well. According to the present invention, the depot formulation can continue to maintain the plasma drug concentration of entecavir over more than several weeks after administration of a single dose, and can dramatically increase patient′s medication compliance.

Description

[0001] The present invention relates to a depot preparation for entecavir fatty acid esters analogs containing an entecavir fatty acid ester derivative,

The present invention relates to a depot preparation for injection administration comprising an entecavir fatty acid ester derivative, and is intended to prepare depot preparations for intramuscular or subcutaneous administration by prodrug-fixing entecavir which has been used only for oral administration .

Entecavir, i.e., the compound 2-amino-9 - [(1S, 3R, 4S) -4-hydroxy-3- (hydroxymethyl) -2- methylcyclopentyl] - purin-6 (9H) -one is a nucleoside-type antiviral agent.

[Chemical Formula 1]

Entecavir is a drug that exhibits the best anti-HBV activity among commercially available anti-HBV (hepatitis B virus) drugs. It is known that the anti-HBV activity of entecavir is 100-fold and 30-fold higher than lamivudine and adefovir dipivoxil, respectively. In addition, there are few side effects, and it shows therapeutic effect in patients showing resistance to lamivudine. Therefore, entecavir has been used as a therapeutic agent for hepatitis B virus.

The treatment of HBV infection requires long-term administration of entecavir, which requires daily oral dosing to avoid the adverse effects of the drug. Therefore, development of a drug capable of exhibiting long-term anti-HBV activity through the administration of a single drug is required. For many reasons, it is desirable to administer entecavir as a sustained or delayed release (depot) formulation that is effective over a prolonged period of time, preferably over about 3 weeks, particularly about 1 month.

Thus, a sustained-release depot based on biodegradable polymer-based microspheres or lipid materials has been attempted. However, due to the low affinity of entecavir for hydrophobic polymers and lipid materials, and high solubility for water-soluble media, there is a problem that drugs loaded in these formulations are not released and are rapidly released into the medium. In addition, there is a problem in that the drug is inevitably lost in the process of loading the drug on the carrier of the polymer or the lipid material, and when the carrier is partially destroyed or deformed, the drug loaded on the inside can be rapidly released.

Korean Patent Publication No. 10-2013-0100464 (published Sep. 11, 2013)

It is an object of the present invention to provide a depot preparation for injection administration comprising an entecavir fatty acid ester derivative, which can be obtained by prodrugating entecavir, which has been used only for oral administration, into a depot formulation for intramuscular or subcutaneous injection administration, To provide a method.

In order to attain the above object, the present invention provides a pharmaceutical composition comprising (a) an entecavir fatty acid ester derivative represented by the following formula (2) having an average particle size of 1 to 50 탆, a pharmaceutically acceptable salt thereof or a hydrate thereof; And (b) a pharmaceutically acceptable carrier.

(2)

Wherein R < ¹ > is ^one of C4 to C20 linear or branched alkyl, alkene or alkane.

In addition, the present invention provides a method for preparing a pharmaceutical composition comprising a first step of dissolving or evenly dispersing an entecavir fatty acid ester derivative, a pharmaceutically acceptable salt thereof or a hydrate thereof in a solvent; A second step of dissolving at least one additive selected from the group consisting of a suspending agent, a surfactant, an isotonizing agent, a buffering agent and a pH adjuster in an anti-solvent; And a third step of adding the solution of the first step to the solution of the second step followed by evaporating the solvent.

The present invention relates to a depot preparation for injection administration comprising an entecavir fatty acid ester derivative, and is intended to prepare depot preparations for intramuscular or subcutaneous administration by prodrug-fixing entecavir which has been used only for oral administration . In order to design an injectable formulation that maintains the effective blood concentration of entecavir for up to three weeks or more, for example up to four weeks after administration of the entecavir fatty acid ester derivative, a fine particle size within the range of 1 to 50 mu m was essential , And that the smaller the particle size of the entecavir fatty acid ester derivative suspension particles, the shorter the time to maintain the blood concentration. Therefore, the depot preparation according to the present invention can exhibit a continuous effect of maintaining the effective blood concentration of entecavir for several weeks or more after single administration, and can greatly improve the patient's compliance with the medication.

Figure 1 shows the blood entecavir drug concentration profile.

The present inventors have found that the entecavir oral therapy conventionally solves the problem that the dietary state greatly affects the rate of absorption of the drug and provides a therapeutically effective sustained-release injection of entecavir for at least 3 weeks or more, , Invented a method for synthesizing fatty acid ester derivatives of entecavir. Entecavir had a solubility of 1700 μg / ml in aqueous solution, while fatty acid ester derivative of entecavir had a low water solubility of less than 100 μg / ml. Considering such low water solubility, when entecavir fatty acid ester derivatives are administered in solid, i.e., in suspension, as muscle or subcutaneous injections, they are present in a solid state in vivo and slowly dissolve and hydrolyse, And it is expected that the release pattern of entecavir will be secured. However, when the synthetic entecavir fatty acid ester derivative is added to the injection water for injection, the wettability is so low that it is not dispersed at all in the injection water and coagulation occurs, which makes it difficult to inject the injected muscle or subcutis. Thus, the active ingredient (entecavir fatty acid ester derivative) is suspended in a pharmaceutically acceptable carrier, and the suspended entecavir fatty acid ester derivative has an average particle size of 1 to 50 占 퐉. The formulation was completed.

(A) an entecavir fatty acid ester derivative represented by the following formula (2) having an average particle size of 1 to 50 mu m, a pharmaceutically acceptable salt thereof or a hydrate thereof; And (b) a pharmaceutically acceptable carrier.

(2)

In the above formula, R ¹ may be any of C4 to C20 linear or branched alkyl, alkene or alkane.

Preferably, the entecavir fatty acid ester derivative represented by the general formula (2) may be entecavir palmitic acid with R ¹ being C15, but is not limited thereto.

Preferably, the pharmaceutically acceptable carrier is water, but is not limited thereto.

Preferably, the entecavir fatty acid ester derivative, a pharmaceutically acceptable salt thereof, or a hydrate thereof is contained in an amount of 0.1 to 30 parts by weight, preferably 0.2 to 15 parts by weight, as an entecavir based on 100 parts by weight of the total amount of the depot preparation But is not limited thereto.

In addition, the present invention may include not only the entecavir fatty acid ester derivative compound of Formula 2, but also possible solvates, hydrates, isomers and the like which can be prepared therefrom.

The pharmaceutically acceptable salts may be in the form of the hydrochloride, bromate, sulfate, phosphate, nitrate, citrate, acetate, lactate, tartrate, maleate, gluconate, succinate, formate, trifluoroacetate, oxalate , A sodium salt, a potassium salt, a lithium salt, a calcium salt, and a magnesium salt may be selected from the group consisting of an alkali metal salt, a sulfonic acid salt, a sulfonic acid salt, a fumaric acid salt, a methanesulfonic acid salt, a benzenesulfonic acid salt, a para toluenesulfonic acid salt,

Microparticle size within the range of 1 to 50 [mu] m is essential to design an injectable formulation that maintains the effective blood concentration of entecavir for up to three weeks or more, such as up to four weeks after administration of an entabeled fatty acid derivative . It has been found that the smaller the particle size of the entecavir fatty acid ester derivative suspended particles, the shorter the time to maintain the blood concentration. Therefore, the present invention preferably has a particle size of 1 mu m or more. On the other hand, with a size of more than 50 microns, the viscosity increases exponentially and is difficult to remove from the vial and involves the difficulty of injecting through a microneedle (e.g., 21 G to 23 G needle).

The present invention has developed and developed a sustained release or sustained release (depot) formulation for a therapeutically effective entecavir fatty acid ester derivative for a period of at least 3 weeks or more, especially about 1 month or more. The expression " at least three weeks or more effective " means that the plasma concentration of the active ingredient entecavir (hydrolyzed from the entecavir fatty acid ester derivative) should be at least about 0.3 ng / ml, the minimum maintenance concentration of oral therapy. On the other hand, in order for the preparation to be " effective ", the plasma concentration should always be maintained below the concentration of approximately 30 ng / ml. If the plasma concentration is higher than the plasma concentration, the patient may experience undesirable side effects or, conversely, below the mean plasma concentration, the systemic resistance of the test agent may be manifested.

&Quot; Fine particle size " means the average volume diameter when measured by laser-light scattering (LLS) techniques. The particle size distribution is measured by the LLS technique and the fine particle size is calculated from the particle size distribution.

The depot preparation of the present invention may further comprise at least one additive selected from the group consisting of a suspending agent, a surfactant, an isotonizing agent, a buffering agent, and a pH adjuster.

In the case of the suspending agent contained in the depot preparation of the present invention, the size of the injected suspended particles is closely related to the maximum blood concentration of the drug and the maintenance period of the effective blood concentration. The size and shape of the suspended particles determine the surface area of the particles. The surface area of the particles controls the dissolution rate of the drug as shown in the Noyes-whitney equation. The smaller the particle size, the wider the surface area, and thus the release rate of the drug increases. Most of the suspension injections, including entecavir fatty acid ester derivatives, exist in the form of a solid state in vivo and dissolve in the body, that is, the process of hydrolysis after elution, and the distribution of the active ingredient (entecavir) , Metabolism, and excretion, the dissolution rate of the drug is a step of determining the maximum blood concentration of the drug and the maintenance period of the effective blood concentration. Therefore, the size control of the suspended particles is a very important factor for devising a therapeutically effective injectable formulation of entecavir for 3 weeks or more, especially about 1 month or more.

Suitable stabilizers, such as suspending agents or surfactants, included in the depot formulations of the present invention include cellulose derivatives such as methylcellulose, sodium carboxymethylcellulose and hydroxypropylmethylcellulose, polyvinylpyrrolidone, alginate Polyoxyethylene derivatives of sorbitan esters such as polysorbate 20 and polysorbate 80, lecithin, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters, Ethylene- and sodium deoxycholate. to be. Preferably, polyethylene glycols or polysorbate 20 are used alone or in combination at a concentration of 0.01 to 10%, more preferably 0.1 to 5% (w / v).

Isotonic agents included in the depot preparation of the present invention include, for example, sodium chloride, dextrose, mannitol, sorbitol, lactose, and sodium sulfate. The suspension conveniently contains 0.1 to 10% (w / v) isotonic agent. Preferably, sodium chloride is used at a concentration of 0.2 to 1.5%.

The buffering agents included in the depot formulations of the present invention include disodium hydrogen phosphate (anhydrous), typically about 0.9% (w / v), to ensure the isotonicity, neutral pH of the solution and the dispersion stability of the suspended entecavir fatty acid ester derivative )) And sodium dihydrogen phosphate monohydrate (typically about 0.6% (w / v)).

The pH adjusting agent contained in the depot preparation of the present invention may be hydrochloric acid or sodium hydroxide, but is not limited thereto.

The depot preparation of the present invention may further contain a preservative and the preservative may be selected from the group consisting of benzoic acid, benzyl alcohol, butylated hydroxyanisole, butylated hydroxytoluene, chlorbutol, gallate, hydroxybenzoate, EDTA, phenol, Is an antimicrobial agent and an antioxidant which can be selected from the group consisting of cresol, metacresol, benztonium chloride, myristyl-gamma-picolinium chloride, phenylmerceric acetate and timmerol. In particular, it is a benzyl alcohol which can be used at a concentration of 2% (w / v) or less, preferably 1.5% (w / v) or less.

In the case of the injection depot preparation of the present invention, securing ease of administration is also important. Preferably, the ability to inject a therapeutically effective amount using a microneedle at as short a time as possible is a basic requirement for injectables. More specifically, all depot formulations should be easily able to be taken into the syringe (e.g., from a vial) and injected through a micro needle (e.g., G23 G needle at 21).

In addition, the injection depot preparation of the present invention can be stored by freeze-drying, and can be resuspended with water for injection immediately before injection.

In addition, the present invention provides a method for preparing a pharmaceutical composition comprising a first step of dissolving or evenly dispersing an entecavir fatty acid ester derivative, a pharmaceutically acceptable salt thereof or a hydrate thereof in a solvent; A second step of dissolving at least one additive selected from the group consisting of a suspending agent, a surfactant, an isotonizing agent, a buffering agent and a pH adjuster in an anti-solvent; And a third step of adding the solution of the first step to the solution of the second step followed by evaporating the solvent.

Preferably, the solvent and the anti-solvent may be in a volume ratio of 1: 0.25 to 2, but are not limited thereto.

Preferably, the entecavir fatty acid ester derivative, a pharmaceutically acceptable salt thereof, or a hydrate thereof is contained in an amount of 0.1 to 30 parts by weight, preferably 0.2 to 15 parts by weight, as an entecavir based on 100 parts by weight of the total amount of the depot preparation But is not limited thereto.

The entecavir fatty acid ester derivatives of the present invention are easily hydrolyzed in the body after being injected into muscles or subcutis and converted to parent drug (entecavir). Therefore, it can be effectively used as a medicine in treating chronic hepatitis B like entecavir.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

<Examples>

100 mg (Example 5), 125 mg (Example 6) and 250 mg (Example 7) of entecavir palmitic acid were dissolved or evenly dispersed in 10 ml of ethanol, and polyethylene glycol 4000 and Tween (Na ₂ HPO ₄ -7H ₂ O) and sodium dihydrogen phosphate dihydrate (NaH ₂ PO ₄ -2H ₂ O) as a buffer were dissolved in 100 ml of water for injection. 10 ml (Example 2), 6 ml (Example 3), 4 ml (Example 4), and 2.5 ml (Example 1), which were maintained at 40 degrees Celsius with ethanol containing entecavir palmitic acid 5-7), and ethanol was evaporated through nitrogen purge (7.5 L / min) to prepare particles. The composition thus prepared is shown in Table 1 below (unit: wt%).

Components (% by weight) Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Example 7 Entecavir palmitic acid (%) 0.39 0.77 1.21 1.91 3.57 4.66 8.69 Polyethylene glycol 4000 (%) 0.2 0.97 4.55 0.96 4.29 0 0 Tween 20 (%) 0.1 0.48 2.27 0 0 0.75 3.47 Sodium chloride (%) 0.88 0.87 0.82 0.86 0.8 0.84 0.78 Sodium dihydrogen phosphate monohydrate (%) 0.19 0.1 0.06 0.19 0.59 0.18 0.06 Disodium hydrogen phosphate (%) 0.45 0.22 0.14 0.44 1.38 0.43 0.14 Hydrochloric acid or
Sodium hydroxide (%) q.s. q.s. q.s. q.s. q.s. q.s. q.s. Injection water (%) q.s. to 100 q.s. to 100 q.s. to 100 q.s. to 100 q.s. to 100 q.s. to 100 q.s. to 100 pH 7.1 7.2 7 7.1 4.79 6.4 7.9

The particle size distribution was measured using a masterizer (MS 2000 (Hydro 2000S), Malvern). The particle size of the examples is shown in Table 2 (unit: mu m).

Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Example 7 D (0.1) 1.98 2.96 2.38 7.92 1.13 1.68 2.54 D (0.5) 2.43 7.56 25.43 43.63 5.25 8.49 14.95 D (0.9) 4.42 14.47 42.17 79.53 11.26 19.87 29.65

<Comparative Example>

80 mg of entecavir palmitic acid was dissolved in 10 ml of ethanol, and 1,000 mg of polyethylene glycol 4000 and 500 mg of Tween 20, 900 mg of sodium chloride as an isotonic agent, 230 mg of disodium hydrogenphosphate (Na ₂ HPO ₄ -7H ₂ O) 99 mg of sodium dihydrogen phosphate dihydrate (NaH ₂ PO ₄ -2H ₂ O) was dissolved in 100 ml of water for injection. Ethanol was added dropwise to 40 ml of injecting water (Comparative Example 1) and 2.5 ml (Comparative Example 2) maintained at 40 degrees of ethanol containing entecavir palmitic acid, and the ethanol was evaporated through nitrogen purge (7.5 L / min) to prepare particles . The composition thus prepared is shown in Table 3 below (unit: wt%).

Comparative Example 1 Comparative Example 2 Entecavir palmitic acid 0.19 3.02 Polyethylene glycol 4000 0.97 0.94 Tween 20 0.49 0.47 Sodium chloride 0.88 0.85 Sodium dihydrogenphosphate dihydrate 0.05 0.09 Sodium dihydrogenphosphate 0.11 0.22 Hydrochloric acid or sodium hydroxide q.s. q.s. pH 0.19 3.02

The particle size distribution was measured using a masterizer (MS 2000 (Hydro 2000S), Malvern). The particle size of the comparative example is shown in Table 4 (unit: mu m).

Comparative Example 1 Comparative Example 2 D (0.1) 0.27 27.98 D (0.5) 0.87 112.67 D (0.9) 1.78 235.24

<Experimental Example 1>

Each of Example 1, Example 2, and Comparative Example 1 was subcutaneously administered to the upper part of four neck beagle dogs with 0.47 mg / kg of body weight using a 23G BD needle. There was no problem in injecting. Blood samples were collected for one month to determine entecavir concentration in plasma. The following pharmacodynamic parameters were calculated from the experimental data (mean ± SD). As can be seen from the plasma concentration profile in plasma according to time, it can be seen that the concentration of 0.3 ng / ml or more can be maintained in Examples 1 and 2 immediately after administration and can be maintained at 0.3 ng / ml or more even after 4 weeks of administration . The peak plasma concentration was also adjusted to below 10 ng / ml. On the other hand, in the case of Comparative Example 1, it is seen that the particle size is small and the faster and higher the highest blood concentration is reached. However, since the disappearance rate is so fast that no drug is detected after 2 weeks, the effective period of treatment is less than 2 weeks (Fig. 1). On the other hand, the pharmacokinetic parameters are shown in Table 5.

Example 1 Example 2 Comparative Example 1 AUC _(0-28day) (ng · h / ml) 1075 ± 215 1067 ± 198 986 ± 112 Cmax (ng / ml) 7.2 ± 3.5 3.5 ± 2.1 16.5 ± 2.9 Tmax (days) 3.2 ± 1.8 6 ± 2.3 1.8 ± 1.0 21 day concentration (ng / ml) 0.5 ± 0.2 0.8 ± 0.4 N / A 28 day concentration (ng / ml) 0.4 ± 0.1 0.5 ± 0.1 N / A

<Experimental Example 2>

The ease of injection for Examples 1-4 and Comparative Examples 1 and 2 was evaluated. Each 10 ml of each formulation was placed in a vial and sampled with a 22G needle attached syringe, and the ease of sampling was evaluated. Thereafter, the injectability of the injected rat was evaluated within 30 seconds (Table 6).

Example 1 Example 2 Example 3 Example 4 Comparative Example 1 Comparative Example 2 Ease of sampling Dragon Dragon Dragon Dragon Dragon Difficult to sample Ease of injection Dragon Dragon Dragon Dragon Dragon Injection difficulty

In Comparative Example 2, the particle size was too large and the viscosity was very large. It was difficult to sample and inject from the vial.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. It is to be understood that various modifications and changes may be made without departing from the scope of the appended claims.

Claims (14)

(a) an entecavir fatty acid ester derivative represented by the following formula (2) having an average particle size of 1 to 50 탆, a pharmaceutically acceptable salt thereof or a hydrate thereof; And
(b) a pharmaceutically acceptable carrier.
(2)

Wherein R &lt; ¹ &gt; is ^one of C4 to C20 linear or branched alkyl, alkene or alkane. The method of claim 1, wherein the depot formulation for injection administration of entecavir fatty acid ester derivative represented by the formula (2) is R ¹ as C15, characterized in that the entecavir palmitate. The composition of claim 1 wherein said pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, bromate, sulfate, phosphate, nitrate, citrate, acetate, lactate, tartrate, maleate, gluconate, succinate, formate, trifluoroacetic acid Is selected from the group consisting of salts, oxalates, fumarates, methanesulfonates, benzenesulfonates, para-toluenesulfonates, camphorsulfonates, sodium salts, potassium salts, lithium salts, calcium salts and magnesium salts. &Lt; / RTI &gt; The depot preparation for injection according to claim 1, wherein the pharmaceutically acceptable carrier is water. The depot preparation for injection according to claim 1, wherein the depot preparation further comprises at least one additive selected from the group consisting of a suspending agent, a surfactant, an isotonizing agent, a buffering agent and a pH controller. 6. The depot preparation for injection according to claim 5, wherein the suspending agent is polyethylene glycol. 6. The depot preparation for injection according to claim 5, wherein the surfactant is polysorbate. 6. The depot preparation for injection administration according to claim 5, wherein the isotonic agent is sodium chloride. The depot preparation for injection according to claim 5, wherein the buffering agent is disodium hydrogenphosphate, sodium dihydrogenphosphate dihydrate or a mixture thereof. The depot preparation for injection according to claim 5, wherein the pH adjusting agent is hydrochloric acid or sodium hydroxide. The enteric-coated depot preparation according to claim 1, wherein the entecavir fatty acid ester derivative, its pharmaceutically acceptable salt or hydrate thereof is contained in an amount of 0.1 to 30 parts by weight as an entecavir relative to 100 parts by weight of the total amount of the depot preparation. . A first step of uniformly dispersing an entecavir fatty acid ester derivative, a pharmaceutically acceptable salt thereof or a hydrate thereof in a solvent;
A second step of dissolving at least one additive selected from the group consisting of a suspending agent, a surfactant, an isotonizing agent, a buffering agent and a pH adjuster in an anti-solvent; And
And a third step of adding the solution of the first step to the solution of the second step and then evaporating the solvent. 13. The method of claim 12, wherein the solvent and the anti-solvent are in a volume ratio of 1: 0.25-2. 13. The method according to claim 12, wherein the entecavir fatty acid ester derivative, its pharmaceutically acceptable salt or hydrate thereof is contained in an amount of 0.1 to 30 parts by weight as an entecavir to 100 parts by weight of the total amount of the depot preparation.

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