TWI459976B - Solidified oil bodies, the producing method, and the use thereof in powder and tablet dosage - Google Patents

Description

Cured oil body, its preparation method and application thereof to powder and tablet dosage form

The present invention relates to a method for curing natural and artificial oil bodies, and more particularly to mixing a mannitol as an excipient with 70% by weight or more and an oil body solution, followed by a drying process to prepare a curing oil. The method of body. The invention also relates to cured curative bodies produced and their use in the preparation of powder or lozenge formulations.

In addition to some basic proteins, plant seed cells also express some unique proteins to provide their physiological needs. In addition to the organelles that basically maintain cell activity, seed cells also have a large number of energy storage devices, such as proteins. Protein body, oil body, starch granule, etc., for the energy source needed for seed germination and post-emergence growth.

These organelles associated with seed germination contain many seed-specific proteins, such as storage proteins in protein bodies and oil membrane proteins (oleosin) in oil bodies. Because the oil film protein itself has a total charge and is negatively charged, it generates an electrostatic repulsion to the oil body, prevents oil body fusion, maintains the integrity and stability of the oil body in the seed cells, and thus stores in the seed for a long time. .

In addition to oil body membrane proteins, it is known that at least two oil body microproteins can bind calcium ions and steroids, respectively, so they are named caleosin and steroleosin. Wherein the oil body calpain has a molecular weight of 27 kDa (see Chen et al., Plant Cell Physiol. 40(10): 1079-1086, 1999).

The inventors of the present invention have confirmed that the three basic components of the oil body: triglyceride, phospholipid and oil body protein (oil body membrane protein or oil body calcium protein), according to their respective proportions in the oil body, The artificial oil body can be successfully formed in a test tube by providing the energy required for oil body reorganization by ultrasonic vibration (Tzen et al., J Biochem 123 : 318-323, 1998; Chen et al., J Agric Food Chem 52). :3982-3987, 2004). Artificial oil bodies are known to be useful as oral delivery systems for anaerobic drugs, and to immobilize enzymes by attaching specific enzymes to oil body proteins on the surface of artificial oil bodies (Chen et al., Biotechnol . Prog . 21 : 1297-130, 2005; Chiang et al, Escherichia coli . Protein Expr . Purif . 52 : 14-18, 2007; Chiang et al, J. Agric . Food Chem . 58 : 11695-11702, 2010).

The artificial oil body must be subjected to a drying process when it is used as a solid dosage form carrier. However, an oil body derived from natural sesame seeds is known to be coalesced during the drying process generally used for the production of foods. And the formation of large oil droplets. Therefore, in order to apply natural and artificial oil bodies to the preparation of solid dosage forms, it is necessary to overcome the problems associated with coalescence during the curing process. Accordingly, the present invention provides a method for curing natural and artificial oil bodies, so that the oil body can maintain similar size and structural integrity before and after drying, and oil body coalescence does not occur, so that it can It is effective for the preparation of oral solid dosage forms such as powder or lozenge formulations.

Accordingly, one aspect of the invention relates to a method of producing a cured oil body powder comprising: (1) providing a natural or artificial oil body solution; (2) adding mannitol to the oil body solution and mixing The final concentration of mannitol is 70% w/v or more; and (3) the resulting mixture is subjected to a drying procedure to obtain an oil body powder in a solidified form. In a particular embodiment of the invention, the final concentration of mannitol in the mixture is from 70% w/v to 90% w/v. In another embodiment of the invention, the final concentration of mannitol in the mixture is 80% w/v.

In a specific embodiment of the invention, the drying procedure is freeze drying. In a specific embodiment of the present invention, the oil body which can be cured by the method of the present invention may be an oil body separated from natural oil or an artificial oil body with or without an active material.

One aspect of the invention relates to a cured oil body powder produced by the oil body solidification method of the present invention. In one embodiment of the invention, the cured oil body powders produced by the present invention are useful in the preparation of solid dosage forms such as powders or lozenges. In another embodiment of the invention, the cured oil body powders prepared in accordance with the invention are useful in carrier compositions for use as excipients for the preparation of pharmaceutical or food compositions.

In view of the various embodiments of the present invention, the various instruments, devices, methods, and related results described below are not intended to be limited to the scope of the present invention.

The other features and advantages of the present invention are further exemplified and illustrated in the following examples, which are intended to be illustrative only and not to limit the scope of the invention.

Example 1: Preparation of solidified oil body powder and its tablet Solidification of natural and artificial oil bodies

Natural sesame seed oil bodies were purified according to the method disclosed by Tzen et al. ( J. Biochem . 121 :762-768, 1997). Caleosin, which is derived from sesame seeds, is prepared according to the method of Liu et al. ( J. Agric . Food Chem . 57 : 2308-2313, 2009).

Add 100, 300, 500, 700 or 900 mg of mannitol to 1 mL of purified sesame oil solution to a final concentration of 10%, 30%, 50%, 70% or 90% (weight/volume) And mix the two together. The resulting mixture was subjected to a freeze-drying procedure. After drying for 24 hours, a solid oil body was obtained as a white powder (Fig. 1).

The oil bodies before and after curing were suspended in 10 mM sodium phosphate buffer (pH 7.5), and placed under a Nikon type-104 optical microscope to observe changes in the appearance of the oil body at a magnification of 400 times. An optical micrograph from the right column of Figure 1 shows that the final concentration is increased by 70% w/v or more. The oil body of the sugar alcohol can effectively prevent the coalescence phenomenon from occurring in the drying process, that is, the solidified oil body powder does not have large oil droplets.

Preparation of solidified oil body lozenges

As described above, the sesame seed oil body solution was dried in the presence of 80% by weight/volume of mannitol to obtain a powdery solidified oil body. Take 100 mg of the solidified oil body powder, mix well with an equal amount of mannitol (100 mg), place the mixture on a Rimek-I rotary tableting machine, and add 10 pressure seconds at 500 lbs using an oblate die. A 200 mg tablet was prepared. The appearance of the cured oil body powder before and after the preparation of the tablet was observed by an optical microscope.

The observation by the optical microscope of Fig. 2 shows that the appearance of the sesame seed oil body before and after curing is consistent in the presence of 80% w/v mannitol, indicating that no oil droplet coalescence occurs, and the above results Match. Moreover, the solidified oil body powder retains the size and structural integrity of the oil body after being formed into a tablet, indicating that the oil body solution is cured by the method of the present invention, thereby preventing not only the occurrence of oil droplet coalescence but also its The stable structure allows it to maintain structural integrity during compression into tablets.

Example 2: Preparation of artificial oil body coated with curcumin

Artificial oil body was prepared from 50 mg of sesame oil (containing 20% PEG 400 and curcumin), 150 μg of diolein phospholipid choline and 250 μg of recombinant oil body troponin in 1 mL of 10 mM sodium phosphate buffer (pH 7.5). As described below. First, the chlorinated phospholipid choline dissolved in chloroform was placed in the bottom of the microcentrifuge tube and placed in a chemical extraction cabinet overnight to evaporate the solvent. After volatilization, sesame oil (containing 20% PEG 400 and curcumin) and recombinant oil body sialoprotein suspended in 1 mL of 10 mM sodium phosphate buffer (pH 7.5) were added to the microcentrifuge tube, followed by an ultrasonic sample processor. Sonics & Materials VCX750, using a 3-mm-diaeter probe (provided with 30% amplification for 20 seconds) for ultrasonic oscillation, then the sample was placed on ice for 5 minutes, and the shock was repeated under the above conditions. Twice, and it has to be wrapped Artificial oil body of curcumin. The purpose of adding 20% PEG 400 is to stabilize the curcumin in sesame oil to provide a lipid matrix as a subsequent artificial oil body.

A stabilized artificial oil body in the form of a yellow emulsified solution has been successfully produced, and the resulting artificial oil body is smaller than the natural sesame seed oil body isolated in the foregoing Example 1 (compare the photographs before the oil body solidification in Figs. 2 and 3). The artificial oil body solution was cured according to the oil body solidification method described in the foregoing Example 1, and the solidified oil body powder was pressed into a tablet in the same manner as described in the examples.

As shown in Fig. 3, the artificial oil body coated with curcumin obtained a yellow powder after the curing drying process. The change in the appearance of the oil body was observed by a microscope, and it was confirmed that the artificial oil body coated with curcumin maintained the structural integrity of the oil body after being cured and formed into a tablet. Therefore, the oil body solidification method of the present invention can be applied to natural oil bodies, and artificial oil bodies with or without active materials.

Example 3: Stability test of cured artificial oil body coated with curcumin

To further evaluate the applicability of cured oil bodies in the manufacture of dosage forms, an accelerated stress test is used (see, Larry, L. Augsburger and Stephen W. Hoag. Pharmaceutical Dosage Forms: Tablets third edition Volume 1: Unit Operation and Mechanical Properties . 2008 , 499) to analyze the stability of solidified oil bodies under long-term storage.

The cured artificial oil body powder and lozenge prepared by the foregoing examples were placed in an oven set to a humidity of 75% at 40 ° C for four months. Thereafter, according to the method of the foregoing Example 1, the appearance of the oil body before and after the four-month storage period was observed by observing the solidified oil body powder and the tablet by an optical microscope. As shown in Fig. 3, after the accelerated stress test, the oil body particles dissolved in the solidified oil body powder or tablet still maintain the integrity of the oil body structure stably, and no obvious deliquescence or dissolution occurs. It is indicated that the solidified oil body obtained by the curing method of the present invention has an excellent potential for use in the preparation of solid dosage forms, including powders or tablets, as a stable pharmaceutical or food composition excipient.

Other specific aspects

All of the features disclosed in this specification can be combined in any combination. Thus, the individual features disclosed in this specification can be replaced by alternative features that are the same, equivalent, or similar. Therefore, the various features disclosed are merely examples of a series of equivalents or similar features, unless otherwise clearly indicated.

From the foregoing description, those skilled in the art can readily determine the essential features of the invention, and various changes and modifications of the invention can be made to adapt to various different uses and conditions without departing from the scope thereof. Therefore, other specific aspects are included in the scope of patent application.

Figure 1 shows the obtained cured sesame oil in the presence of different concentrations (10% w/v, 30% w/v, 50% w/v, 70% w/v and 90% w/v) of mannitol A photograph of the appearance of the powder (left panel); and a photomicrograph of the oil body sample (right panel) after 30 minutes of suspension in 10 mM sodium phosphate buffer (pH 7.5) was observed under an optical microscope. The ruler line segment in the figure represents 10 μm.

Figure 2 is a photograph of the appearance of a cured natural sesame oil in powder and its compacted tablet form (top panel) and photomicrograph (below). The cured sesame oil powder in the figure was obtained by mixing an oil body solution with 80% by weight/volume mannitol and drying. The optical micrographs were obtained by observing the oil bodies in the oil body powder or tablets before and after curing in a 10 mM sodium phosphate buffer (pH 7.5) for 30 minutes and then taking samples. The ruler line segment in the figure represents 10 μm.

Figure 3 is a photograph (top panel) and a photomicrograph (pictured below) of a cured artificial oil body in powder form (containing curcumin) and its compressed tablet form. The cured artificial oil body powder in the figure is obtained by mixing an oil body solution with 80% by weight/volume mannitol and performing a drying process. Optical micrographs will be before, after, and after 4-month storage. The oil body in the oil powder or tablet was suspended in 10 mM sodium phosphate buffer (pH 7.5) for 30 minutes, and then sampled and observed. The ruler line segment in the figure represents 10 μm.

Claims (12)

A method for producing a solidified oil body, comprising: (1) providing a natural or artificial oil body solution; (2) adding mannitol to the oil body solution and mixing to make mannitol The final concentration is 70% by weight/volume or more; and (3) the resulting mixture is subjected to a freeze-drying procedure to obtain an oil body powder in a cured form. The method of claim 1, wherein the final concentration of the mannitol is from 70% by weight to 90% by weight/volume. The method of claim 1, wherein the final concentration of the mannitol is 80% w/v. The method of claim 1, wherein the oil body is an oil body of natural origin. The method of claim 1, wherein the oil body is an artificial oil body. The method of claim 6, wherein the artificial oil body further comprises an active material. A solidified oil body powder obtained by the method of claim 1 of the patent application. A solidified oil body powder as described in claim 7 for the preparation of a solid dosage form. The solidified oil body powder according to claim 8, wherein the solid dosage form is a tablet. A carrier for preparing a pharmaceutical or food composition, which comprises The solidified oil body powder of item 7 of the benefit range. The carrier of claim 10, wherein the pharmaceutical or food composition is formulated as an oral solid dosage form. The carrier according to claim 11, wherein the oral solid dosage form is a tablet.