Patent Application
20210338588
The present invention relates to the pharmaceutical field, and specifically, to a lyophilized preparation of BPA and a preparation method.
Boron neutron capture therapy (BNCT) is using boron (10B)-containing drugs having high capture cross section characteristics for thermal neutrons, to produce two heavy charged particles, 4He and 7Li, through 10B(n,α)7Li neutron capture and a nuclear fission reaction. The two charged particles have average energy of about 2.33 MeV and characteristics of high linear energy transfer (LET) and a short range. The LET and the range of α particles are 150 keV/μm and 8 μm respectively, while the LET and the range of 7Li heavy charged particles are 175 keV/μm and 5 μm respectively. A total range of the two particles is approximately equivalent to a size of a cell, and therefore, radiation damage to organisms can be limited to a cellular level. When the boron-containing drugs are selectively aggregated in tumor cells, the tumor cells can be locally killed by selecting an appropriate neutron source, without causing too much damage to normal tissues.
4-Boronophenylalanine (BPA) is a 10B-containing drug containing that is studied frequently at present. Because it is difficult to dissolve BPA in water, addition of a solubilizer is generally needed. In addition, the BPA is dissolved under the action of a strong acid or base, and then, the resultant solution is regulated to approximately the physiological pH value, and after being prepared into a sterile injection through sterilization, is applied to patients or animals. The solution needs to be prepared while being used. The cumbersome preparation process causes extremely inconvenient clinical use of the drug, and sterility cannot be guaranteed, which limits application thereof. The Patent CN103100094B discloses a freeze-drying process applicable to L-BPA, where after L-BPA and a solubilizer, fructose, are mixed, the L-BPA is dissolved, and is freeze-dried under vacuum for 22-26 hours.
Therefore, it is necessary to propose a new technical solution to resolve the foregoing problem.
To resolve the foregoing problem, an aspect of the present disclosure provides a method for preparing a lyophilized preparation of BPA and a BPA preparation prepared by using the method, where the preparation prepared by using the method has good stability and a small content of impurities.
The method for preparing a lyophilized preparation of BPA includes a solution preparation process and a freeze-drying process, where the solution preparation process includes: (1) dissolving BPA and polyol in an aqueous solution by using a base to obtain a clear solution; (2) regulating the clear solution back to 7.5<pH≤8.5 by using an acid, to obtain a BPA solution; and the freeze-drying process includes: (3) subpackaging the BPA solution and freeze-drying under a condition with a vacuum of 10-20 Pa, to obtain the lyophilized preparation.
Implementations of this aspect may include one or more of the following features.
In another preferred embodiment, the temperature of the solution in the solution preparation process is controlled to be lower than or equal to 60° C., preferably, 18 to 50° C., and more preferably, 18 to 40° C.
In another preferred embodiment, a ratio of parts by weight the BPA to the polyol is 1:1-1.3, preferably, 1:1.1-1.25.
In another preferred embodiment, a vacuum of the freeze-drying is 10-20 Pa, preferably, 10-11 Pa.
In another preferred embodiment, the clear solution is regulated back to a pH value of 7.6-8.1, to obtain a BPA solution.
In another preferred embodiment, the base includes: lithium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, and magnesium hydroxide.
In another preferred embodiment, in the solution preparation process, the BPA and the polyol are dissolved in the aqueous solution by using the base, to obtain a clear solution, and a pH value of the clear solution is 8.5-9.5.
In another preferred embodiment, the polyol includes: fructose, lactose, sorbitol, maltose, mannitol, xylitol, ribose, glucose, and sucrose.
In another preferred embodiment, after the solution preparation process, and before the freeze-drying process, the method further includes filtering the BPA solution in a filtering step.
In another preferred embodiment, a time of the freeze-drying process is 39-80 hours.
In another preferred embodiment, a vacuum in the freeze-drying process is 10-11 Pa.
In another preferred embodiment, the freeze-drying process further includes pre-freezing the BPA solution in a pre-freezing process. Preferably, the temperature of the pre-freezing process is −20° C. to −50° C. Preferably, a time of the pre-freezing process is 5-15 hours.
In another preferred embodiment, the freeze-drying process further includes subliming the BPA solution in a sublimation process. Preferably, the temperature of the sublimation process is −20° C. to −35° C. Preferably, a vacuum of the sublimation process is 10-20 Pa. Preferably, a time of the sublimation process is 30-55 hours.
In another preferred embodiment, the freeze-drying process further includes desorption drying the BPA solution in a desorption drying process. Preferably, the temperature of the desorption drying process is 20° C. to 40° C. Preferably, a vacuum of the desorption drying process is 10-20 Pa. Preferably, a time of the desorption drying process is 4-10 hours.
The second aspect of the present disclosure provides a lyophilized preparation of BPA prepared by using the method according to the first aspect of the present invention.
The third aspect of the present disclosure provides a composition including the lyophilized preparation of BPA according to the second aspect of the present disclosure.
The fourth aspect of the present disclosure provides a kit including the lyophilized preparation of BPA according to the second aspect of the present disclosure.
Further areas of applicability will become apparent from the description provided herein. It should be understood that the description and specific examples are intended for purposes of illustration only and are not intended to limit the scope of the present disclosure.
The accompanying drawings illustrate one or more embodiments of the disclosure and together with the written description, serve to explain the principles of the disclosure. Wherever possible, the same reference numbers are used throughout the drawings to refer to the same or like elements of an embodiment.
Unless otherwise defined, all technical and scientific terms in this specification have the same meanings as that usually understood by a person skilled in the art to which the claimed subject belongs. Unless otherwise specified, all patents, patent applications, and publications cited in this specification are incorporated herein by reference in their entirety.
It should be understood that the above brief description and the following detailed description are exemplary and only used for explanation, and are not intended to limit the subject of the present invention. In this application, unless otherwise specified, the plural forms are included when the singular form is used. It should be noted that, unless otherwise clearly specified in this specification, the singular form used in this specification and claims includes the plural referents. It is also noted that, unless otherwise specified, the use of “or”, “alternatively” means “and/or”. In addition, the terms “comprise”, “include”, and other grammatical forms such as “comprising” and “including” are not limiting. Section titles in this specification are only used for the purpose of organizing the text, and should not be explained as limitations to the subject. All documents or parts of a document cited in this application, including but not limited to patents, patent applications, articles, books, operating manuals, and papers, are incorporated herein by reference in their entirety.
The “base” described in the present invention is mainly inorganic base, including, but not limited to, lithium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, and magnesium hydroxide. The “acid” described in the present invention includes inorganic acid and organic acid. The inorganic acid, for example, includes, but is not limited to, perchloric acid, hydroiodic acid, sulfuric acid, hydrobromic acid, hydrochloric acid, nitric acid, iodic acid, oxalic acid (oxalic acid), sulfurous acid, phosphoric acid, pyruvic acid, carbonic acid, citric acid, hydrofluoric acid, malic acid, gluconic acid, formic acid, lactic acid, benzoic acid, acrylic acid, ethylic acid (acetic acid), propionic acid, stearic acid, hydrosulfuric acid, hypochlorous acid, boric acid, trifluoroacetic acid, methanesulfonic acid, benzenesulfonic acid. The preferred acid is hydrochloric acid.
The “BPA” described in the present invention is 4-dihydroxyboryl phenylalanine, of which a chemical formula is
where B includes 10B and 11B. When being 10B, B may be used as a drug for the BNCT, and includes two isomers, respectively,
Both configurations should be included in the scope of protection of the present invention, and the BPA of the present invention is preferably L-BPA.
Method for Preparing a Lyophilized Preparation of BPA
The present invention provides a method for preparing a lyophilized preparation of BPA, including a solution preparation process and a freeze-drying process, where the solution preparation process includes: (1) dissolving BPA and polyol in an aqueous solution by using a base to obtain a clear solution; (2) regulating the clear solution back to 7.5<pH≤8.5 by using an acid, to obtain a BPA solution; and the freeze-drying process includes: (3) subpackaging the BPA solution and freeze-drying under a condition with a vacuum of 10-20 Pa, to obtain the lyophilized preparation. To ensure that chemical properties of components in the solution do not change, in the solution preparation process, the temperature of the solution is controlled to be lower than or equal to 60° C. The “solution” mentioned herein refers to any of the “solutions” in the solution preparation process. Preferably, the temperature is 18 to 50° C., and more preferably, 18 to 40° C.
A ratio of parts by weight of the BPA to polyol affect dissolution of the BPA, but the ratio of parts by weight is not specially limited. Preferably, a ratio of parts by weight the BPA to the polyol is 1:1-1.3, preferably, 1:1.1-1.25. The polyol includes: fructose, lactose, sorbitol, maltose, mannitol, xylitol, ribose, glucose, and sucrose.
In the solution preparation process, the BPA and the polyol are dissolved in the aqueous solution by using the base, to obtain a clear solution, and a pH value of the clear solution is 8.5-9.5. Further, the clear solution needs to be regulated back to a pH value of 7.6-8.1, to obtain a BPA solution.
In the freeze-drying process, a vacuum of the freeze-drying is 10-20 Pa, preferably, 10-11 Pa. A time of the freeze-drying process is 39-80 hours. During actual operation, the above may be determined according to actual parameters in the solution preparation process, and are not specially limited. The freeze-drying process includes: a pre-freezing process, a sublimation process, and a desorption drying process. Parameters, such as a temperature, a time, and a vacuum, in the three processes may be independently selected according to actual needs. In a preferred solution, the BPA solution is freeze-dried according to the following conditions, to obtain a lyophilized preparation of BPA: in a pre-freezing process: reducing the temperature of a sample to −20° C. to −60° C., and maintaining for 5-15 hours to completely freeze the sample; in a sublimation process: raising the temperature to −15° C. to −35° C., maintaining a vacuum of 10-11 Pa, and maintaining for 30-55 hours; and in a desorption drying process: heating a partition plate to raise the temperature to 0° C. to 40° C., maintaining for 4-10 hours, and maintaining a vacuum of 10-11 Pa in the entire desorption drying process. In a preferred solution, in the pre-freezing process, the temperature sometimes may be regulated a plurality of times according to actual needs, for example, is first reduced to −60° C., and maintained for a period of time, and then, is raised to −20 to −50° C. In a preferred solution, in the desorption drying process, the temperature may be first raised to a relatively low temperature, and then be gradually raised to a relatively high temperature, for example, the temperature is first raised to 0° C. and maintained for a period of time, and then, is raised to 30±10° C.
Mainly Advantages of the Present Invention Include:
Through wide and thorough researches, the inventor developed a lyophilized preparation of BPA and a preparation method, including a solution preparation process and a freeze-drying process. In the freeze-drying process, optimized process parameters are used, to greatly prevent incomplete removal of moisture in the freeze-drying process and poor temperature control from exerting adverse impact on the product form, so that preparation has good stability and a low content of impurities. The present invention is made on such basis.
The following further describes the present invention with reference to the specific embodiments. It should be understood that the following descriptions are only optimal implementations of the present invention, and should not be regarded as limitations to the protection scope of the present invention. On the basis of a full understanding of the present invention, the experimental methods without specific conditions in the following embodiments are usually in accordance with the conventional conditions or in accordance with the conditions recommended by the manufacturer. A person skilled in the art may make non-essential changes to the technical solutions of the present invention, and such changes should be included in the protection scope of the present invention. Unless otherwise specified, the percentage and the parts are the percentage by weight and the parts by weight respectively.
(1) Add 1.0 kg of BPA, 1.1-1.3 kg of fructose, and an appropriate amount of water for injection into a solution preparation tank, wash containers containing the BPA and the fructose with water 3 times, and transfer the water used for washing into the solution preparation tank, and stir for 10 min.
(2) Add a sodium hydroxide solution into the solution preparation tank to dissolve the BPA, and stir until the solution is clear.
(3) Add a hydrochloric acid solution, to regulate a pH value of the solution to 7.6.
Whether the solution after being regulated back and prepared by using the hydrochloric acid is examined. The hydrochloric acid is added into the solution, to regulate the solution to a different pH value close to the physiological pH value. Experiment results indicating whether precipitates are generated in the solution under the condition of pH=7.3-8.5 within 48 hours are observed, as shown in Table 1. In addition, the pH values under the conditions of pH=7.6, 8.0, and 8.5 respectively are shown in Table 2, and the pH value of the solution does not change significantly within 24 hours.
The BPA solution is freeze-dried according to the following conditions, to obtain a lyophilized preparation of BPA:
As shown in Table 3, the lyophilized sample is redissolved with water to prepare a solution, the solution stands at room temperature for 24 hours, and none of measurement results of the pH value, the clarity, the transmittance, and the content significantly changes. After the present product is prepared and then, stands at room temperature for 24 hours, the solution has good stability.
As shown in
The process is described in detail as follows.
{circle around (1)} Preparation procedure: adding water for injection into a solution preparation tank, adding prescription doses of BPA and an excipient, adding a sodium hydroxide solution, washing containers containing the excipient and the sodium hydroxide solution respectively with water for injection, and transferring the water used for washing to the solution preparation tank; stirring until the solution is clear; and regulating the pH value, adding the remaining water for injection, stirring evenly, detecting an intermediate, and filtering, where the excipient is the polyol described in the present invention.
{circle around (2)} Filling procedure: regulating a filling volume according to a content of the intermediate, filling, partially adding stoppers, and feeding into a freeze-drying box.
{circle around (3)} Freeze-drying procedure: including: a pre-freezing stage, a sublimation stage, and a desorption drying stage.
{circle around (4)} Unboxing and capping procedure: unboxing: protecting semi-finished products under a 100-level laminar flow, and removing and scrapping an unqualified product that, for example, lacks a stopper, has a displaced stopper, or has a broken bottle; and capping: sampling before, during, and after production to detect appearances of samples.
{circle around (5)} Packaging procedure: labeling, boxing, encasing, and storing.
Although the illustrative embodiments of the present invention have been described above in order to enable those skilled in the art to understand the present invention, it should be understood that the present invention is not to be limited the scope of the embodiments. For those skilled in the art, as long as various changes are within the spirit and scope as defined in the present invention and the appended claims, these changes are obvious and within the scope of protection claimed by the present invention.
| Number | Date | Country | Kind |
|---|---|---|---|
| 201910341883.5 | Apr 2019 | CN | national |
This application is a continuation application of International Application No. PCT/CN2020/086745, filed on Apr. 24, 2020, which claims priority to Chinese Patent Application No. 201910341883.5, filed on Apr. 26, 2019, the disclosures of which are hereby incorporated by reference.
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/CN2020/086745 | Apr 2020 | US |
| Child | 17375071 | US |
