JP2020200295A - Lyophilized preparation - Google Patents

Abstract

To provide a lyophilized preparation that contains an optically active trans-astaxanthin derivative having excellent antioxidant activity, and excellent water solubility and the like, a salt thereof, or a high-purity derivative thereof or a salt thereof.SOLUTION: A lyophilized preparation contains an optically active trans-astaxanthin derivative represented by formula (I), a salt thereof, or high-purity compounds thereof (where m1, m2, n1 and n2 are the same or different and represent an integer of 1-6).SELECTED DRAWING: None

Description

The present invention relates to a lyophilized preparation containing a trans-astaxanthin derivative.

Astaxanthin is a pigment substance contained in some algae, krill, shrimp, sea bream, salmon, etc., has strong antioxidant activity, and acts as a factor that protects the living body from ultraviolet rays and lipid peroxidation. It is believed that. As a compound that improves the water solubility and oral absorption of astaxanthin and has an excellent anti-inflammatory effect, various functional groups are additionally introduced into the hydroxy group of the cyclohexenone ring at both ends of astaxanthin in Patent Document 1 as follows (I). ), An optically active trans-astaxanthin derivative has been reported.

(In the equation, m ₁ , m ₂ , n ₁ and n ₂ mean the same or different integers 1 to 6, respectively.)

International Publication No. 2019/6674

However, the compound of the formula (I) is not always sufficient in terms of stability, and is unstable due to decomposition by hydrolysis or the like especially in an aqueous solution, and the compound of the formula (I) is stably contained for a long period of time. Securing storable compounds was a major issue.
An object of the present invention is to provide a long-term storable preparation which stably contains an optically active trans-astaxanthin derivative of the formula (I), a salt thereof and a high-purity compound thereof.

Therefore, as a result of diligent studies to obtain a long-term storable preparation containing the optically active trans-astaxanthin derivative of the formula (I) stably, the present inventors have made a result of diligent studies to obtain the derivative of the formula (I), a salt thereof or a high purity. A lyophilized preparation containing these compounds was found, and the present invention was completed.

That is, the present invention provides the following inventions [1] to [10].

[1] A lyophilized preparation containing an optically active trans-astaxanthin derivative represented by the formula (I), a salt thereof, or a high-purity compound thereof.

(In the equation, m ₁ , m ₂ , n ₁ and n ₂ mean the same or different integers 1 to 6, respectively.)
[2] The lyophilized preparation according to [1], wherein m ₁ and m ₂ in the formula (I) mean an integer of 1 respectively, and n ₁ and n ₂ mean an integer of 3 respectively.
[3] The lyophilized preparation according to [1] or [2], wherein the salt is a sodium salt.
[4] The lyophilized preparation according to any one of [1] to [3], which further contains a saccharide.

[5] The lyophilized preparation according to [4], wherein the saccharide is lactose, trehalose or a mixture thereof.
[6] The lyophilized preparation according to [4] or [5], which further contains a pH adjuster.
[7] The lyophilized preparation according to [6], wherein the pH adjuster is a phosphate buffer, a citrate buffer or a mixture thereof.
[8] The lyophilized preparation according to [6] or [7], wherein the pH adjusting agent is a pH adjusting agent that adjusts the pH to a neutral range.
[9] The lyophilized preparation according to any one of [1] to [8], wherein the lyophilized preparation is a lyophilized powder.
[10] The lyophilized preparation according to any one of [5] to [9], wherein the lyophilized preparation is a lyophilized pharmaceutical preparation.

The lyophilized preparation of the present invention is excellent as a long-term storable preparation in which an optically active trans-astaxanthin derivative represented by the formula (I), a salt thereof or a high-purity compound thereof exists in a stable state. It is extremely excellent as a medicinal preparation of the compound of formula (I), which is an antioxidant having excellent antioxidant activity, water solubility and orally absorbable.

The optically active trans-astaxanthin derivative used in the present invention is represented by the above formula (I).
In the compound of the formula (I) of the present invention, the trans is basically the isomer of the trans, that is, the double bond portion capable of forming the geometric isomer in the intermediate carbon chain portion in the astaxanthin basic skeleton. It means that it is a body (hereinafter referred to as all trans isomers).
Further, in the compound of the formula (I), the optical activity means that the two ester groups bonded to the astaxanthin basic skeleton both have an S configuration with respect to the basic skeleton. Therefore, the configuration of the asymmetric carbon atom in the cyclohexenone skeleton of the formula (I) is 3S, 3'S.

Among the compounds of the above formula (I), compounds in which m ₁ and m ₂ mean an integer of ₁ and n ₁ and n ₂ each mean an integer of 3, and salts thereof are preferable.

The compound of formula (I) can form a pharmaceutically acceptable salt by subjecting a basic substance or a basic compound desired because it has a carboxyl group in the molecule to a normal salt forming reaction. Such salts include, for example, alkali metal salts such as sodium salts, potassium salts, lithium salts; alkaline earth metal salts such as calcium salts, magnesium salts, lysine salts, ornithine salts, amino acids such as arginine salts. Salts can be mentioned, with sodium salts being preferred.

The optically active trans-astaxanthin derivative of the formula (I) having high purity means that the trans-astaxanthin derivative having high purity of at least 95% or more, preferably 98% or more of the active ingredient is present. The salt of this compound can be obtained as a salt of the highly pure optically active trans-astaxanthin derivative of the formula (I) according to the present invention by subjecting it to a normal salt forming reaction.

The high-purity optically active trans-astaxanthin derivative of the present invention and a salt thereof can be used for the treatment of diseases involving active oxygen whose effects have been confirmed as free forms of astaxanthin. Diseases involving active oxygen that can be improved / prevented include, for example, hyperlipidemia, obesity, glucose intolerance, hypertension, insulin resistance, metabolic syndrome, fatty liver, diabetes, and diabetic complications (eg, retina). Diseases, nephropathy, neuropathy, cataracts, diabetic luteal edema, coronary artery disease, etc.), steatosis, non-alcoholic steatosis (NASH), hepatitis C, arteriosclerosis, gestational diabetes, polycystic ovary syndrome, heart Vascular disease (eg, ischemic heart disease), atherosclerosis, vascular insufficiency, stroke (eg, cerebral thrombosis, cerebral embolism, transient cerebral ischemic attack, cerebral bleeding, submucosal bleeding), gout, inflammation Sexual disorders (eg, pain, fever, rheumatoid arthritis, inflammatory enteritis, acne, sunburn, psoriasis, eczema, allergic disorders, GI ulcers, malaise, autoimmune disorders, pancreatitis, etc.), osteoarthritis, gastric ulcers , Cancer, osteoporosis, cataract, glaucoma, eye fatigue, dementia (eg, Levy body dementia, cerebrovascular dementia, frontotemporal dementia, juvenile dementia, alcoholic dementia, etc.), depression Disease, bipolar disorder, schizophrenia, chronic fatigue, disused muscle atrophy, muscle atrophy, sarcopenia, malaise, uric acid inhibitory effect, hair growth promoting effect, wound improvement, lung disease (eg, chronic obstructive pulmonary disease) , Asthma), pain disorders (eg, inflammatory pain, neuropathy pain, fibromyalgia, cancerous pain, migraine, etc.), urinary disorders (eg, interstitial cystitis, overactive bladder, etc.) and surgery Later postoperative pain and improvement of wound healing can be mentioned.
Preferably, it is preferably used as a preventive agent and / or a therapeutic agent for enteritis, particularly preferably colitis, especially ulcerative colitis and knee osteoarthritis.

In a lyophilized preparation containing an optically active trans-astaxanthin derivative represented by the formula (I) of the present invention, a salt thereof or a high-purity compound thereof, the optically active trans-astaxanthin derivative, a salt thereof or a compound thereof having high purity. The amount of the compound is not particularly limited, and it may be blended within a range in which the compound can be solubilized in the manufacturing process of this preparation.

In this lyophilized preparation, sugars, specifically lactose, trehalose, and mixtures thereof can be preferably used as excipients, and lactose is particularly preferable. The amount used thereof is not particularly limited, but usually, from the viewpoint of long-term storage stability of the compound of the formula (I), the mass of the optically active trans-astaxanthin derivative of the formula (I) of high purity or a salt thereof (1). 1 part by mass to 100 parts by mass, preferably 5 parts by mass to 10 parts by mass, and particularly preferably 8 parts by mass to 9 parts by mass.

Regarding the pH of this lyophilized preparation, from the viewpoint of long-term storage stability of the compound of the formula (I), it is in the neutral range, specifically, 6.3 to 7.4, particularly preferably 6.8 to. It may be adjusted to about 7.1.
For pH adjustment, sodium hydrogen phosphate, potassium dihydrogen phosphate, a phosphate buffer in which they are mixed, trisodium citrate, citric acid and a citric acid buffer in which citric acids and their citric acids are mixed can be preferably used. it can.

Next, a method for producing the lyophilized preparation of the present invention will be described.
The optically active trans-astaxanthin derivative of formula (I) or a salt thereof is usually suspended or dissolved in distilled water for injection, and an L-lysine solution, an aqueous sodium hydroxide solution, or a mixture thereof is added as necessary. Mix to prepare an aqueous solution of the compound.
An aqueous solution of the above pH adjusting agent may be added to the aqueous solution to adjust the pH to the desired pH, and then filtration may be performed using a filter, and if necessary, aseptic filtration treatment may be performed to obtain an aqueous solution before freezing. In the preparation of the above aqueous solution, it is also possible to appropriately bubbling using an inert gas such as nitrogen gas, if necessary.

Further, in the preparation of the above-mentioned aqueous solution before freezing, additives such as a surfactant and a preservative may be added as needed, and the amount of the addition may be an amount usually used.

An aqueous solution of the optically active trans-astaxanthin derivative of the formula (I) or a salt thereof obtained as described above is dispensed into a vial, an ampoule or the like as necessary, and then by using a usual freeze-drying technique. The desired freeze-dried preparation of the present invention can be produced.

The lyophilized preparation of the present invention means a vial, an ampoule, or the lyophilized powder itself, which contains or fills the lyophilized powder obtained after lyophilization in a vial or ampoule, and they are collectively lyophilized. It can be a formulation.
The obtained lyophilized powder can be applied and developed in various pharmaceutical administration preparations such as capsule filling as needed.

Hereinafter, the present invention will be described in detail with reference to Reference Examples, Examples and Test Examples, but the present invention is not limited thereto.
The API used in the examples is the following compound.
4- (3- {4- [18- (4 (S)-[3- (3-carboxylpropyl) ureidoacetoxy] -2,6,6-trimethyl-3-oxycloheyxa-1-enyl) -3,7,12 , 16-tetramethyloctadeca-1,3,5,7,9,11,13,15,17-nonaenyl] -3,5,5-trimethyl-2-oxycloheyxa-3-enyl-1 (S) -oxycarbonylmethyl} ureido) butyric acid

Examples (Table 1. Prescription No. 1)
60.914 g of API was suspended in 2100 mL of water for injection and bubbling with nitrogen gas while heating. To the suspension, 3000 mL of an L-lysine solution (prepared by adding 37.7 g of L-lysine hydrochloride and 100 mL of an aqueous sodium hydroxide solution to 5 L of water for injection) was added little by little to dissolve the suspension. After confirming that the pH of the solution was 7.6 to 7.9, the solution was bubbled with nitrogen gas. 540 g of lactose hydrate was added to the solution, and the mixture was stirred and dissolved. 200 mL of phosphate buffer (prepared by dissolving 29.88 g of sodium hydrogen phosphate hydrate and 14.40 g of potassium dihydrogen phosphate in water for injection and measuring up to 300 mL) was added to the solution, and the pH of the solution was added. Was confirmed to be around 6.9. The weight of the solution was adjusted to 6222 g with water for injection, and then pressure filtration was performed with nitrogen gas using a filter having a pore size of 0.22 μm and a membrane material, a polyether sulfone (PES). The filtrate was dispensed and filled into a vial with a filling amount of 2.70 (± 0.02) mL. After filling, the rubber stopper was half-stopped and lyophilized to obtain a lyophilized preparation vial.
Similarly, the prescription No. 2 to 13 lyophilized preparations were obtained.

Test Examples The lyophilized preparations obtained in Examples were subjected to stability tests.
Specifically, the lyophilized preparation of the present invention is prepared under the conditions of a so-called accelerated stability test (40 ° C., 75 ± 5% relative humidity storage) and a harsh stability test (60 ° C., no particular conditions other than temperature). After storage for a specific period, the content of the drug substance was measured according to the following quantification method. The residual rate of the drug substance was calculated with the quantitative value at the start of storage as 100% and shown in Tables 2 and 3.

Comparative test example (stability of the above drug substance in aqueous solution)
The residual rate of the drug substance is calculated with the quantitative value at the start of storage of the redissolved solution as 100%.

From the results of Tables 2 and 3 and the results of Table 4, the lyophilized preparation of the present invention improved the stability of the compound of the formula (I) and was excellent as a long-term storage preparation of the compound of the formula (I). It was confirmed that there was. As the excipient, lactose and trehalose are preferable, and lactose is particularly preferable. Further, it can be seen that the pH is preferably in the neutral range, more preferably pH 6.3 to 7.4, and even more preferably pH 6.8 to 7.1.

(API quantification method)
The lyophilized preparation (containing 27 mg of API ⁾ was dissolved in ¹ mL of distilled water and solution ^{Note 1)} to make 50 mL. Internal standard solution ^{Note 2)} 5 mL was added to 5 mL of the solution to make 50 mL of the solution, which was used as a sample solution. Separately, about 50 mg of the standard API product was dissolved in 10 mL of distilled water, 5 mL of L-lysine solution ^{Note 3)} , and solution ^{Note 1)} to make 100 mL. 5 mL of the internal standard solution was added to 5 mL of the solution to make 50 mL of the solution, which was used as a standard solution.
Note 1) 0.60 g of anhydrous sodium dihydrogen phosphate and 0.71 g of anhydrous disodium hydrogen phosphate were dissolved in distilled water to make 1000 mL. 500 mL of acetonitrile for HPLC was added to 500 mL of the solution.
Note 2) Methyl red sodium solution (1 → 2000)
Note 3) About 75 mg of L-lysine was dissolved in distilled water to make 20 mL.
For the sample solution and standard solution 10 [mu] L, were tested by liquid chromatography, and measuring the area ratio Q _T and Q _S of the peak area and API internal standard, in accordance with the following formula, it was calculated content of API.

For the measurement of liquid chromatography, a column filled with octadecylsilylated silica gel for liquid chromatography of 5 μm in a stainless steel tube having an inner diameter of 4.6 mm and a length of 15 cm was used. As the mobile phase, a 0.025% trifluoroacetic acid acetonitrile solution (mobile phase A) and a 0.025% trifluoroacetic acid aqueous solution (mobile phase B) were used, and the solutions were fed according to the concentration gradient shown in Table 5. The measurement wavelength was 474 nm, the column temperature was around 25 ° C., and the flow rate was 1.0 mL / min.

Claims (10)

A lyophilized preparation containing an optically active trans-astaxanthin derivative represented by the formula (I), a salt thereof, or a high-purity compound thereof.

(In the equation, m ₁ , m ₂ , n ₁ and n ₂ mean the same or different integers 1-6.) The lyophilized preparation according to claim 1, wherein m ₁ and m ₂ in the formula (I) mean an integer of 1 respectively, and n ₁ and n ₂ mean an integer of 3 respectively. The lyophilized preparation according to claim 1 or 2, wherein the salt is a sodium salt. The lyophilized preparation according to any one of claims 1 to 3, further comprising a saccharide. The lyophilized preparation according to claim 4, wherein the saccharide is lactose, trehalose or a mixture thereof. The lyophilized preparation according to claim 4 or 5, further comprising a pH adjuster. The lyophilized preparation according to claim 6, wherein the pH adjuster is a phosphate buffer, a citric acid buffer, or a mixture thereof. The lyophilized preparation according to claim 6 or 7, wherein the pH adjusting agent is a pH adjusting agent that adjusts the pH to a neutral range. The lyophilized preparation according to any one of claims 1 to 8, wherein the lyophilized preparation is a lyophilized powder. The lyophilized preparation according to any one of claims 5 to 9, wherein the lyophilized preparation is a lyophilized pharmaceutical preparation.