CN1264501C

CN1264501C - Process for preparing white arsenic nano particles

Info

Publication number: CN1264501C
Application number: CN 200410041822
Authority: CN
Inventors: 王子妤; 张东生; 路新丽
Original assignee: Southeast University
Current assignee: Southeast University
Priority date: 2004-08-31
Filing date: 2004-08-31
Publication date: 2006-07-19
Anticipated expiration: 2024-08-31
Also published as: CN1615904A

Abstract

The present invention relates to a technology for preparing medicine arsenic nanoparticles for treating tumors. As2O3 and hydrochloric acid (HCl) are mixed by a proportion of mass of each milligram to a volume of each milliliter of 1: (0.02-0.2), and magnetically stirred for 10 to 20 minutes so as to be fully mixed, dissolved and added with an ethanol by a volume proportion of hydrochloric acid to ethanol of 1: (5-10) to be stirred for 10 to 20 minutes; ultrasonic treatment is carried out for 10 to 20 minutes at a temperature of 50 DEG C to 60 DEG C; distilled water is added by a volume ratio of ethanol to distilled water of 1: (4 to 5) to be fully stirred for 10 to 20 minutes; subpackage is carried out, and sterile As2O3 nanoparticles which can be used for treating tumors are obtained after a process of degerming through millipore filtration. As2O3 nanoparticles prepared by adopting the technology of the present invention present is colorless liquid through the observation of the unaided eyes, and the density of the As2O3 nanoparticles is uniform, and the form of As2O3 nanoparticles is presented as a circular or elliptical shape through the observation by an electron microscope. The present invention has the advantages of good dispersivity, and grain diameter of about 80 nm and 40 nm.

Description

Preparation process of arsenic nano granule

Technical Field

The present invention relates to a new medicine preparation of traditional medicine arsenic for curing tumor, namely a preparation process of nano granules.

Background

Arsenic trioxide (As)₂O₃) Has a long history of being used for treating leukemia, As in 1999₂O₃The medicine is approved by the national drug administration, passes the FDA approval in 25.9.2000 months, is widely applied to clinic and achieves remarkable effect. A large number of studies have shown that As₂O₃The mechanisms of antitumor are mainly growth inhibition, differentiation induction and apoptosis. As₂O₃The powder is insoluble in water and produces significant toxic side effects if administered orally. The dosage form used clinically is As₂O₃The injection mainly contains arsenous acid, and after intravenous administration, due to the fact that the arsenic concentration in blood plasma is rapidly increased and rapidly diffused to surrounding tissues, the injection can cause patients to have digestive tract symptoms, peripheral neuritis, dryskin, pigmentation, even toxic and side effects such as renal dysfunction or hydrothorax and ascites, and particularly epidemiological investigation finds that the occurrence of certain tumors is related to the accumulation amount of arsenic. We are therefore seeing As₂O₃The clinical application value of the composition is realized, and simultaneously, the composition can be used for preparing a new medicament form, so that the dosage is reduced, the curative effect is improved, and the composition has important clinical significance. Nanotechnology is considered to be one of the most promising fields in the 21 st century, and in the field of drug research, due to the continuous penetration and influence of nanotechnology, a profound revolution in the field of drugs is initiated, and a new term of nano drugs appears. The nano-drug is nano-particles (NP) orNanospheres (NS), Nanocapsules (NC), etc. are carriers that are combined with drugs in a manner to produce drugs that may have particle sizes in excess of 100nm, but are typically less than 500 nm. The nano-drug can also be a nano-particle prepared by directly processing the raw material drug. The National Science and Technology Committee (NSTC) proposed nanotechnology development report forecast in 2000: about 50% of the drugs in the market sales will be applied by nanotechnology in the whole world in the next 10 years. In recent years, nanotechnology has begun to be widely regarded in traditional Chinese medicine research and application in China, and a new concept of nano traditional Chinese medicine is created, so that certain progress has been made in the aspects of traditional Chinese medicine effective components, effective parts, raw medicines, compound compounds, novel preparations and the like manufactured by adopting nanotechnology. Xuhuibi and the like prepare nano abalone shell and realgar particles, and research shows that the nano abalone shell and realgar particles have good anti-tumor effect.

Disclosure of Invention

The invention provides a process for preparing nano-scale arsenic nanoparticles, which can improve the drug effect of arsenic on treating solid tumors and reduce the dosage, thereby effectively reducing the toxic and side effects of arsenic.

The invention adopts the following technical scheme:

a preparation method of arsenicum nanoparticle for treating tumor comprises adding As₂O₃Mixing with hydrochloric acid (HCl) at a ratio of 1: 0.02-0.2 in terms of mass per milligram to volume per milliliter, magnetically stirring for 10-20 minutes to fully mix and dissolve the HCl and the HCl, adding ethanol at a ratio of 1: 5-10 in terms of volume of the HCl and the ethanol, continuously stirring for 10-20 minutes, ultrasonically treating for 10-20 minutes at 50-60 ℃, adding distilled water at a ratio of 1: 4-5 in terms of volume of the ethanol and the distilled water, fully stirring for 10-20 minutes, subpackaging, and filtering for sterilization to obtain sterile As for tumor treatment₂O₃And (3) nanoparticles.

The preparation principle of the invention is the following equation: ；；； its structural features and physical characteristics: (1) form; (2) and the particle size; (3) and performing energy spectrum analysis.

Compared with the prior art, the invention has the following advantages:

as produced by the process of the present invention₂O₃The nano-particlemeat is in a colorless liquid state, the density of the nano-particle meat is relatively uniform under an electron microscope, the nano-particle meat is in a round or oval shape, the dispersibility is good, and the particle size is about 80nm and 40 nm. Electron microscope energy spectrum analysis, using a probe to hit on a single nanoparticle, wherein only the components of carbon, oxygen, arsenic and copper (carbon, oxygen and copper are all components of copper net, copper is removed As background in fig. 4) can be hit in the energy spectrum, which indicates that the prepared nanoparticle is indeed As₂O₃. When a material is processed to nanometer size (1-100 nm), it exhibits many specific properties due to its size close to the wavelength of light and its large surface area. Surface effects refer to the smaller the size of the particle, the larger the surface area. E.g. a specific surface area of 90m at a particle size of 10nm²·g^-1: when the particle diameter is 5nm, the specific surface area is 180m²·g^-1(ii) a When the particle diameter is as small as 2nm, the specific surface area is increased to 450m²·g^-1. Such a large specific surface area results in a rapid increase in the surface energy due to the increasing number of atoms present on the surface, and thus the nanoparticles have a high chemical activity. As₂O₃Belongs to mineral traditional Chinese medicine, the powder of the traditional Chinese medicine is insoluble in water, has high electron density under an electron microscope, is in a square, polygonal or irregular crystal shape, and has the average diameter of about 5 mu m. The traditional arsenous acid injection for clinical use is an alkaline solution, although the particle size of the particles in the solution is small, the dispersibility is poor, the particles are easy to form agglomeration, and the particle size of the particles is increased after a plurality of particles are agglomerated. As prepared by the present invention₂O₃The nano-particles have very good dispersibility and little agglomeration among the particles, so the particles have very large sizeSurface energy and chemical activity to make As₂O₃The efficacy of the composition is enhanced.

As prepared by the present invention for the laboratory₂O₃The nanoparticles are used for carrying out in-vitro treatment experiments on human hepatoma cells SMMC-7721, and MTT and flow cytometry experiments prove that As is₂O₃The nanoparticles can generate obvious cytotoxic effect on tumor cells, and the cell survival rate is obviously lower than that of As with the same concentration₂O₃Solution treatment group. As compared with the clinical arsenous acid injection through in vitro cell experiments₂O₃The nanoparticles show better antitumor effect. As₂O₃The nano-particle is expected to become As₂O₃A new dosage form for treating tumor is As₂O₃Opens up a new way for clinical application.

Drawings

FIG. 1 is As₂O₃A projection electron micrograph of the powder;

FIG. 2 is As₂O₃The diameter of the nanoparticle is about 80nm in a transmission electron microscope picture 1 of the nanoparticle;

FIG. 3 is As₂O₃Transmission electron microscope photograph 2 of nanoparticles having a diameter of about 40 nm;

FIG. 4 is As₂O₃The nanoparticles were analyzed by scanning electron microscopy.

FIG. 5 As₂O₃Solution and As₂O₃The comparative survival rate concentration after the nanoparticles are used for treating SMMC-7721 cells is 5 mu mol/L (b) the concentration is 10 mu mol/L, and NP is the nanoparticles.

FIG. 6 As₂O₃Solution and As₂O₃After the nanoparticles are used for treating SMMC-7721 cells, a normal control group of apoptosis rate (A) is determined by a flow cytometer; (B)5 μmol/LAs₂O₃A solution group; (C)5 μmol/LAs₂O₃A set of nanoparticles; (D) 10. mu. mol/LAs₂O₃Solution group: (E) 10. mu. mol/LAs₂O₃Nano particleAnd (4) grouping.

Detailed Description

A preparation method of arsenicum nanoparticle for treating tumor comprises adding As₂O₃Mixing with hydrochloric acid (HCl) at a ratio of 1: 0.02-0.2 in terms of mass per milligram to volume per milliliter, magnetically stirring for 10-20 minutes to fully mix and dissolve the HCl and the HCl, adding ethanol at a ratio of 1: 5-10 in terms of volume of the HCl and the ethanol, continuously stirring for 10-20 minutes, ultrasonically treating for 10-20 minutes at 50-60 ℃, adding distilled water at a ratio of 1: 4-5 in terms of volume of the ethanol and the distilled water, fully stirring for 10-20 minutes, subpackaging, and filtering for sterilization to obtain sterile As for tumor treatment₂O₃Nanoparticles, in this example, As₂O₃The ratio of hydrochloric acid and hydrochloric acid can be selected to be 1: 0.03, 1: 0.05, 1: 0.08, 1: 0.1, 1: 0.13, 1: 0.145, 1: 0.18, 1: 0.19, the volume ratio of hydrochloric acid to ethanol can be selected to be 1: 6, 1: 7.5, 1: 9, the volume ratio of ethanol to distilled water can be selected to be 1: 4.2, 1: 4.5, 1: 4.65, 1: 4.9.

Claims

1. A process for preparing the nano-particles of arsenic trioxide As the medicine to treat tumor₂O₃Mixing with hydrochloric acid at a ratio of 1: 0.02-0.2 in terms of mass per milligram to volume per milliliter, magnetically stirring for 10-20 minutes to fully mix and dissolve the hydrochloric acid, adding ethanol at a ratio of 1: 5-10 in terms of volume of hydrochloric acid to ethanol, continuously stirring for 10-20 minutes, carrying out ultrasonic treatment at 50-60 ℃ for 10-20 minutes, adding distilled water at a ratio of 1: 4-5 in terms of volume of ethanol to distilled water, fully stirring for 10-20 minutes, subpackaging, and filtering and sterilizing by micropores to obtain the sterile As for tumor treatment₂O₃And (3) nanoparticles.