CN100335038C - Preparation process and application of inorganic salt nanometer particle containing biological active material and enveloped Chinese medicinal ingredient - Google Patents
Preparation process and application of inorganic salt nanometer particle containing biological active material and enveloped Chinese medicinal ingredient Download PDFInfo
- Publication number
- CN100335038C CN100335038C CNB2005100411635A CN200510041163A CN100335038C CN 100335038 C CN100335038 C CN 100335038C CN B2005100411635 A CNB2005100411635 A CN B2005100411635A CN 200510041163 A CN200510041163 A CN 200510041163A CN 100335038 C CN100335038 C CN 100335038C
- Authority
- CN
- China
- Prior art keywords
- inorganic salt
- chinese medicine
- bioactive substance
- nano particles
- nanometer particle
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Abstract
The present invention relates to a preparation method of an inorganic salt nanometer particle containing bioactive substances and encapsulated Chinese medicinal components and an application thereof. Firstly, inorganic salt nanometer particle containing bioactive substances are prepared; active substances and bioactive substances of Chinese medicine are encapsulated; the inorganic salt nanometer particles containing bioactive substances and encapsulated Chinese medicine components are acquired. In disease treatment, the inorganic salt nanometer particle is used as a carrier for transferring the bioactive substances or used as a vaccine adjuvant. The present invention has the advantages that because the inorganic salt nanometer particle has the defects of easy adhesion, poor dispersion rate and requirement for improving physicochemical performance, the dispersion rate of the inorganic salt nanometer particle and the encapsulation efficiency of the bioactive substances can be increased after encapsulation by the active substances of the Chinese medicine; the effective components in the Chinese medicine also enable the nanometer particle to be converted into a nanometer carrier with a certain biological function from a pure carrier; simultaneously, the formed nanometer particle can be biologically degraded in vivo. The present invention has simple preparation technology, high stability of a product, minimum difference among different batches and easy industrialization.
Description
Technical field
The present invention is the transmission system of a kind of medicine and vaccine.Especially in the transmission system of medicine or vaccine, provide a kind of and contain bioactive substance and seal the preparation method of the inorganic salt nano particles of Chinese medicine ingredients.
Background technology
Nanotechnology is 21 century one of the revolutionary technology of tool, is a boundary science technology interdisciplinary.It is the crystallization of multi-door subjects such as biology, medical science, materialogy, chemistry, physics.Nanotechnology is as a kind of space platform technology, and its scope has related to many subjects and technical field.Especially aspect medical domain, pharmaceutics development and biological engineering and the medical science equipment revolutionary breakthrough being arranged, become one of focus in medical science, biomedicine and the study of pharmacy.
Nanotechnology relates to the development of the development technology of medical material, Medical Equipment, pharmaceutics in medical research.The preparation of nanoparticle and application then are to have the break-through point of hope in the pharmaceutics and have great market and using value.Nanoparticle typically refers to size particle (flat its energy: " Nano medication and nano-carrier system " in the 100nm-1000nm scope in pharmaceutics research.Chinese Journal of New Drugs 11 the 1st phases of volume of ground in 2002, numbering 1003-3734 (2002) 01-0042-05).
In biomedical sector, utilize nanometer or micron particle as diagnostic tool, improve the sensitivity and the correctness of diagnosis, thereby reach early stage, quick and accurately diagnosis in conjunction with computer technology.Be the healing of tumor disease offer help (Borm PJ, Kreyling W.J Nanosci Nanotechnol.2004 May; 4 (5): 521-31.Toxicological hazards of inhaled nanoparticles--potential implications for drugdelivery).Simultaneously, because the diagnostic kit of nanometer preparation is convenient to storage and transportation, thereby reduced diagnostic fees usefulness.Being particularly suitable for extensive on-the-spot EPDML generaI investigation and backwoodsman hospital uses.
Nanoparticle has following advantage as drug delivery system in materia medica and pharmaceutics:
1, nanoparticle is particularly suitable as various types of pharmaceutical carriers because of its size, physicochemical property and surface characteristic, and forms the transmission system of medicine by different combinations.Because its size belongs to subcellular fraction, sub-micrometer range, in vivo by the efficient of cellular uptake than the big 15-250 of micron particle doubly and can pass inferior mucous layer, and micron only rests on the mucosa top layer.Nanoparticle can also pass blood brain barrier effectively, thereby the transmission medicine enters brain, increases the concentration of medicine at brain, effectively treats brain diseases.For example cerebroma can not well can not get effective treatment (Lisa Brannon-Peppas and James O.Blanchette.Adv Drug DelivRev.2004 Sep 22 by blood brain barrier owing to medicine exactly; 56 (11): 1649-59.Nanoparticle and targeted systems for cancer therapy. and Kreuter J.J Nanosci Nanotechnol.2004 May:4 (5): 484-8.Influence of thesurface properties on nanoparticle-mediated transport of drugs to the brain.).
2, some nanoparticle is can be biodegradable.Carry medicine when nanometer and enter in the body,, guaranteed drug concentrations thereby medicine continues release along with the degraded of nanoparticle, improved the action time of medicine, reduce dosage, alleviated the toxic and side effects of medicine, reduced the discomfort reaction that patient causes because of taking medicine.
3, nanoparticle can directedly be carried medicine, especially can directedly carry antitumor drug, behind intravenous injection nanometer antitumor drug, find that medicine accumulates in around the tumor, improved medicine antitumor characteristic, reduced dosage, can obviously reduce the reaction of patient the toxic and side effects of medicine.The function of this targeted medicine of nanoparticle has improved the medicine local concentration, has increased the curative effect of medicine, be particularly suitable for treatment (the Lockman PR of brain and ocular disease, Mumper RJ, Khan MA, Allen DD.Drug Dev Ind Pharm.2002 Jan; 28 (1): 1-13.Nanoparticle technology for drugdelivery across the blood-brain barrier.).
4, improve the stability of macromolecular drug.Especially peptide class and protein drug.This class medicine is synthesized in the nanoparticle, can protect medicine when oral, not destroyed and protect medicine not by enzymatic degradation (Gavini E by the low pH value environment of gastrointestinal, Chetoni P, Cossu M, Alvarez MG, SaettoneMF, Giunchedi P.Eur J Pharm Biopharm.2004 Mar; 57 (2): 207-12.PLGA microspheres forthe ocular delivery of a peptide drug, vancomycin using emulsification/spray-drying as thepreparation method:in vitro/in vivo studies. and Morishita M, Takayama K.NipponRinsho.2002 Sep:60 Suppl 9:516-21.The design of oral insulin delivery systemsbased on smart hydrogels).
5, discover that recently not only can targeting transmission medicine can also transmit medicine to specific tissue and cell with nanometer as the medicine of carrier arrives specific organelle, further improve bioavailability of medicament (Takeuchi H, Yamamoto H, Kawashima Y.Adv Drug Deliv Rev.2001 Mar23; 47 (1): 39-54.Mucoadhesive nanoparticulate systems for peptide drugdelivery.).
Nanoparticle occupies more and more important position in the development of drug delivery and pharmaceutics and development, the medicine that will become potentialization soon carries system.Especially along with the increase of the demand and the market proportion of biologics, the demand of Nano medication transmission system will improve thereupon.
At present in drug delivery system the nanoparticle of common research have: solid lipid nanoparticle of forming by fatty acid, aliphatic alcohol and phospholipid; Nanosphere or nanometer bundle by bioabsorbable polymer material preparations such as polylactic acid, the poly-third glue fat, poly-chitose, gelatin; The nanoparticle of forming by the copolymer of polylactic acid and Polyethylene Glycol or poly-chitose and derivant thereof.Its synthetic method has polymerization; Supercritical technology; The cohesion dispersion method; The high pressure homogenization method; The fusion dispersion method; Solvent evaporated method and emulsifying/solvent diffusion method.At present China the nanoparticle of normal research lipid nanometer, nanocapsule, nanosphere and Nano capsule are arranged.Wherein liposome nano granule is the most a kind of (flat its energy, " present situation of nano-drug preparation is with following " 2002-09-03, a Chinese Medicine of research both at home and abroad; And Maitra, et al.United State Patent 6,555,376.July 24,2002; And Bell, etal.United State Patent 635,5271.March, 2002; And Fulton et al.United StatePatent 5,266,205.Nov, 1993).
Though synthetic medicine of nanotechnology and vaccine delivery system have very big progress in recent years, but still have some important obstacles, make nanoparticle stagnate aspect the industrialization as drug delivery system.The problem that exists has (Rao GC, Kumar MS, Mathivanan N, Rao ME.Pharmazie.2004 Jan at present; 59 (1): 5-9.Nanosuspensions as the most promising approach innanoparticulate drug delivery systems.):
At first, the raw material of most of nanoparticles is organic compound or derivatives thereofs, the building-up process complexity.Change the structure of macromolecular drug with meeting in the bonded preparation process of macromolecular drug, thereby reduce its activity.For example, PLGA with the bonded process of pharmaceutical grade protein in can lose about 30% protein active.
Secondly, some nanoparticles in vivo can not biodegradation, thereby influences the slow release and the controlled-release function of entrained medicine, and because the gathering in vivo of particle might produce certain side effect.
The 3rd, most of nanoparticle building-up process complexity, raw material mostly is some organic macromolecule materials again, thereby the poor repeatability between each batch product, the size of particle, surperficial physicochemical characteristic and concentration are criticized in difference very big difference between the product, be difficult to the suitability for industrialized production of control nanoparticle, bring very big obstacle for the industrialization of nanoparticle.The liposome nanometer is modal.
The 4th, there is certain side effect in some particle carrier system, and the mechanism of the side effect of its generation it be unclear that.For example, with the carrier of liposome as influenza vaccines, facial paralysis rate occurred frequently appears in clinical report, forbids in the European market at present.
The 5th, the polymer nanoparticle is after repeatedly taking, and is slow because of its biodegradation rate, may bring out the general toxic reaction, and may the toxigenicity metabolite in degradation process.
The 6th, the nanoparticle of macromolecule organic can't carry out autoclave sterilization, can only sterilize with gamma ray.This also has influence on the application of nanoparticle at human body.
The 7th, do not find still that at present a kind of method can produce lipid nanometer particle and effectively controlling cost on a large scale.
The 8th, it is poor that solid lipid nanoparticle and medicine combine rear stability, and medicine very easily decomposes from nanoparticle, thereby influences the envelop rate of medicine.
Still not having a kind of nanometer transmission system so far formally is applied to clinical.
Summary of the invention:
The objective of the invention is to: mostly be a series of problems that solid lipid nanoparticle and nanocapsule, nanosphere and Nano capsule exist in using in preparation with as drug delivery system at the nanoparticle that in materia medica and pharmaceutics, adopts at present, provide a kind of in vivo can biodegradation and contain bioactive substance and seal the preparation method of the inorganic salt nano particles of Chinese medicine ingredients.
The object of the present invention is achieved like this: a kind of preparation method that contains the inorganic salt nano particles that bioactive substance and Chinese medicine ingredients seal is characterized in that:
A) preparation comprises the inorganic salt nano particles of bioactive substance:
After 0.01～1.0 mole saline solution, bioactive substance solution, 0.01～1.0 mole phosphoric acid solution mixed by 1: 1: 1, stirred 48 hours down at 4 ℃, supersound process 15 minutes, under 4 ℃, centrifugal 15 minutes of 8500rpm removes the supernatant that contains free bioactive substance solution, uses the deionized water suspended particles, concentration at 5mg/ml forms the inorganic salt nano particles that contains bioactive substance, and its size dimension is 100nm～800nm;
B) seal the active substance and the bioactive substance of Chinese medicine:
Under the room temperature, after will containing the active substance of inorganic salt particle, Chinese medicine of bioactive substance and bioactive substance and mixing by 1: 10: 10, concentration by 10mg/ml is suspended in pH value 7.0,0.01 in the phosphate buffer of mole, on shaking table, shook under 4 ℃ 2 hours, 4 ℃, centrifugal three times of 8500rpm, each 15 minutes, obtain the inorganic salt nano particles that contains bioactive substance and seal Chinese medicine ingredients;
Described saline solution is 0.01～1.0 mole a manganese chloride with the deionized water preparation, or magnesium chloride, or calcium chloride solution; Described bioactive substance is a protein; The active substance of described Chinese medicine is an astragalus polysaccharides, or Cordyceps polysaccharide, or pachyman, or lycium barbarum polysaccharide.
The inorganic salt nano particles that bioactive substance and Chinese medicine ingredients seal that contains that obtains is stored in 4 ℃ or freeze-dried back.
In the present invention: described protein is HSV-2 protein, or I (chlamydia) protein matter (MOMP), or human growth hormone (hGH).
The invention has the advantages that: because the calcium phosphate or the inorganic salt nano particless such as magnesium phosphate or manganese phosphate that form in preparation process are easy to inter-adhesive, dispersivity, the Surface Physical Chemistry characteristic needs to improve, adopt the inorganic salt nano particles of the active substance parcel back formation of Chinese medicine not only can change ionic surface electrode, improve their dispersion rate, and improved the envelop rate of biological active matter confrontation inorganic salt nano particles; Effective ingredient in the Chinese medicine (compositions such as polysaccharide, saponin) itself has certain physiological function, in case with after inorganic salt nano particles combines, can make nanoparticle become the nanometer portable object with certain biological function from pure portable object; Simultaneously, the nanoparticle of formation biodegradation in vivo.
The present invention be advantageous in that: not only preparation contains bioactive substance and seals the technology of inorganic salt nano particles of Chinese medicine ingredients simple, and the stability of product is high, different batches batch between difference minimum, be easy to industrialization.
Description of drawings
Fig. 1: the illustraton of model that contains the inorganic salt nano particles that bioactive substance and Chinese medicine ingredients seal;
Fig. 2: contain the technological process of the preparation method of the inorganic salt nano particles that bioactive substance and Chinese medicine ingredients seal;
Fig. 3: contain the inorganic salt nano particles stereoscan photograph that bioactive substance and Chinese medicine ingredients are sealed;
Fig. 4: the size distribution that contains the inorganic salt nano particles that bioactive substance and Chinese medicine ingredients seal;
Fig. 5: contain the inorganic salt nano particles that bioactive substance and Chinese medicine ingredients seal and carry the human growth hormone extracorporeal releasing test;
Fig. 6: contain inorganic salt nano particles that bioactive substance and Chinese medicine ingredients seal and carry the content in blood plasma after the administration of human growth hormone via intranasal application;
Fig. 7: the simple viral II type antigen of human body (HSV-2) external from the nanoparticle release test.
In Fig. 1, Fig. 2: 1, inorganic salt nano particles, 2, the active substance of Chinese medicine, 3, bioactive substance, 4, saline solution, 5, phosphoric acid solution, 6, the inorganic salt nano particles that contains bioactive substance, 7, agitator, 8, high speed centrifuge, 9, the ultrasonic disruption instrument, 10, shaking table, 11, freezer dryer.
The specific embodiment
Embodiment 1: contain preparation method and the application thereof of the inorganic salt nano particles that the usefulness human growth hormone (hGH) of Chinese medicine flavone and saponin constituent seals
Preparation before the preparation
Saline solution: with the manganese chloride of 0.01～1.0 mole of deionized water preparation (or magnesium chloride, or calcium chloride solution).
Phosphoric acid solution: with the phosphoric acid solution of 0.01～1.0 mole of deionized water preparation.
Bioactive substance solution: the ratio in every 100ml phosphate buffer+10mg human growth hormone prepares 10% human growth hormone solution.
The active substance solution of Chinese medicine: by the complex active substance solution that is prepared into Chinese medicine of every 100ml normal saline+10mg flavone and saponin.
PH value is that 7.0,0.01 moles phosphate buffer can outsourcing.
Fig. 1 is the illustraton of model that contains bioactive substance and seal the inorganic salt nano particles of Chinese medicine ingredients, as seen from Figure 1, inorganic salt nano particles 1 inside in the model is bioactive substance 3, be encapsulated with the active substance 2 of Chinese medicine in the periphery of the inorganic salt nano particles 1 that contains bioactive substance 3, seal bioactive substance 3 at last again.What present embodiment reflected is the inorganic salt nano particles that contains human growth hormone and seal Chinese medicine ingredients flavone and saponin, in inorganic salt nano particles 1 selection is human growth hormone, what the periphery was selected all is flavone and saponin, the still human growth hormone that outermost layer is sealed.
Fig. 2 contains bioactive substance and seals the technological process that the preparation method of the inorganic salt nano particles of Chinese medicine ingredients relates to, and as seen from Figure 2, preparation process is as follows:
A) preparation comprises the inorganic salt nano particles of bioactive substance:
With 0.01～1.0 mole saline solution 4, bioactive substance 3 solution, 0.01～1.0 mole phosphoric acid solution 5 by mix at 1: 1: 1 place agitator 7 after, stirred 48 hours down at 4 ℃, passed through ultrasonic disruption instrument 9 supersound process 15 minutes again, then under 4 ℃, is centrifugal 15 minutes of 8500rpm through high speed centrifuge 8 at rotating speed, removal contains the supernatant of free bioactive substance solution, use the deionized water suspended particles, concentration at 5mg/ml forms the inorganic salt particle 6 that contains bioactive substance, and its size dimension is 100nm～800nm;
B) seal the active substance and the bioactive substance of Chinese medicine:
Under the room temperature, after will containing the inorganic salt nano particles 6 of bioactive substance, the active substance 2 of Chinese medicine (present embodiment is the active substance of Chinese medicine complex) and bioactive substance 3 and mixing by 1: 10: 10, concentration by 10mg/ml is suspended in pH value 7.0,0.01 in the phosphate buffer of mole, on shaking table 10, shook 2 hours under 4 ℃, then under 4 ℃, is centrifugal three times of 8500rpm through high speed centrifuge 8 at rotating speed, each 15 minutes, obtain the inorganic salt nano particles that contains bioactive substance and seal Chinese medicine ingredients.
The inorganic salt nano particles that contains bioactive substance and seal Chinese medicine ingredients that obtains should be stored in 4 ℃ or by freezer dryer 11 freeze-dried backs.
In concrete preparation process, described saline solution 4 can be selected 0.01～1.0 mole manganese chloride of deionized water preparation for use, or magnesium chloride, or calcium chloride solution; The active substance 2 of described Chinese medicine can be selected the monomer of Chinese medicine or the active substance of complex for use, and they comprise Cordyceps polysaccharide, or pachyman, or astragalus polysaccharides, or glycyrrhizic acid, or lycium barbarum polysaccharide and comprise Cordyceps, or Poria, or the Radix Astragali, or Radix Glycyrrhizae, or the crude extract of Fructus Lycii; Described bioactive substance 3 solution can be selected for use and contain protein, or fat-soluble small-molecule drug, or water miscible small-molecule drug, or the antibody class medicine, or the solution of polypeptide drug, or gene genetic active matter liquid.
After preparation finishes, can calculate the concentration of bioactive substance in the nanoparticle, concrete grammar is: use the deionized water suspended particles, detect the concentration of free bioactive substance, calculate the concentration of bioactive substance in the nanoparticle, detect the size and the distribution of particle with Backman Counter N5 Plus.The inorganic salt nano particles stereoscan photograph that contains bioactive substance and seal Chinese medicine ingredients is referring to Fig. 3, and the size distribution that contains bioactive substance and the inorganic salt nano particles of sealing Chinese medicine ingredients is referring to Fig. 4.
Because the saline solution that adopts is a calcium chloride in the present embodiment, the active substance solution of the Chinese medicine that adopts is Chinese medicine flavone and saponin solution, the bioactive substance solution that adopts is 10% human growth hormone solution, its preparation be the inorganic salt nano particles that a kind of human growth hormone that contains Chinese medicine flavone and saponin is sealed.Obtain the inorganic salt nano particles that a kind of human growth hormone (hGH) that contains Chinese medicine flavone and saponin constituent is sealed.
The inorganic salt nano particles that has carried out containing human growth hormone in the application respectively and sealed Chinese medicine ingredients is carried the experiment of human growth hormone release in vitro and carries after human growth hormone body via intranasal application implements mucosal drug delivery the content of human growth hormone in blood plasma by the inorganic salt nano particles that contains human growth hormone and seal Chinese medicine ingredients.Wherein, the experimental data of human growth hormone release in vitro is referring to Fig. 5; The experimental selection mouse of the content after the administration in blood plasma, the content after the administration in blood plasma is referring to Fig. 6.
Embodiment 2:
Present embodiment prepares the HSV-2 inorganic salt nano particles vaccine that contains the Chinese medicine astragalus crude extract according to method and the step of embodiment 1, the difference of it and embodiment 1 only is: the active substance of the Chinese medicine complex of selection is: the Chinese medicine astragalus crude extract, the bioactive substance of selection is: HSV-2 protein.
The host mainly relies on immune system with partial immunologic function, especially vagina to the resistance of a lot of property disease virus, and main opposing function is played in the invasion of property disease pathogen.Therefore, in the immunity and treatment of property disease, vagina immunization route and vagina administration approach become research emphasis.We synthesize the transmission system of back as the HSV-2 vaccine with the complex of the crude extract of inorganic salt nano particles and Chinese medicine astragalus according to the method for embodiment 1, realize that medicine and vaccine are from the direct administration of vagina.In the experiment, at first carried out the simple viral II type antigen of human body (HSV-2) external from the nanoparticle release test, experimental result is referring to Fig. 7, then, we have selected mouse, are divided into 5 groups, adopt the medicine of following ingredients: 1, blank; 2, Chinese medicine astragalus crude extract; 3, HSV-2 is prepared into vaccine; 4, crude extract in the Chinese medicine astragalus and HSV-2 protein are prepared into vaccine; 5, the inorganic salt nano particles that contains the Chinese medicine astragalus crude extract of the present invention's preparation is the HSV-2 vaccine of transmission system.Grouping is carried out the anti-experiment of HSV-2 Idiotype antibody horizontal (referring to table 3) and host immune protection test (referring to table 4) respectively from the direct administration of vagina.By experiment as can be seen: the present invention can improve the local concentration of medicine, thereby increases drug effect; Simultaneously, improve reproductive system local immunity power, thereby remove the virus of invasion; Avoided the metabolism of liver to medicine.Also can reach whole body therapeutic effect effectively by vagina administration.
Table 3: antibody concentration (ug/ml)
| | Contrast | The Radix Astragali | HSV-2 | The HSV-2+ Radix Astragali | HSV-2+ nanometer+Radix Astragali |
| | 0 | 0 | 10 | 90 | 200 |
| | 0 | 2 | 30 | 200 | 400 |
Table 4: clinical symptoms
| Time behind the viral infection (my god) | Contrast | The Radix Astragali | HSV-2 | HSV-2+Radix Astragali | HSV-2+ nanometer+ |
| 6 | 2.3 | 2 | 1.8 | 1.4 | 1.1 |
| 8 | 8 | 6.2 | 5.8 | 3.3 | 0 |
| 10 | 10.5 | 9 | 7.2 | 4.8 | 0 |
Present embodiment prepares the vagina chlamydia inorganic salt nano particles vaccine that contains Chinese medicine astragalus active polysaccharide material according to method and the step of embodiment 1, the difference of it and embodiment 1 only is: the monomer reactivity material of the Chinese medicine of selection is: Chinese medicine astragalus active polysaccharide material, the bioactive substance of selection is: vagina I (chlamydia) protein matter (MOMP).
We select 25 vagina chlamydia patients at clinical employing, are divided into 5 groups, adopt the medicine of following ingredient: 1, blank; 2, Chinese medicine astragalus polysaccharide; 3, vagina I (chlamydia) protein matter (MOMP) is prepared into vaccine; 4, Chinese medicine astragalus polysaccharide and vagina I (chlamydia) protein matter (MOMP) protein are prepared into vaccine; 5, the inorganic salt nano particles that contains the Chinese medicine astragalus polysaccharide of the present invention's preparation is vagina I (chlamydia) protein matter (MOMP) vaccine of transmission system, respectively from administered intramuscular.Vagina I (chlamydia) protein matter Idiotype interferon concentration level experiment (referring to table 5) and vagina I (chlamydia) protein matter Idiotype antibody concentration level experiment (referring to table 6).By experiment as can be seen: the present invention can improve the antigenicity of vagina I (chlamydia) protein matter, thereby increases vagina chlamydia patient general immunity power; Simultaneously, improve reproductive system local immunity power, thereby remove invading bacteria (referring to table 7).Inorganic salt nano particles vagina I (chlamydia) protein matter (MOMP) vaccine that contains the Chinese medicine astragalus polysaccharide by administered intramuscular also can reach topical therapeutic effect effectively.
Table 5:gamma interferon concentration (pg/ml)
| The immunity back time (week) | Contrast | The Radix Astragali | MOMP | The MOMP+ Radix Astragali | MOMP+ nanometer+ |
| 8 | 0 | 10 | 50 | 140 | 470 |
| 16 | 0 | 10 | 75 | 155 | 920 |
Table 6: antibody concentration (ug/ml)
| | Contrast | The Radix Astragali | MOMP | The MOMP+ Radix Astragali | MOMP+ nanometer+Radix Astragali |
| | 0 | 1 | 12 | 125 | 240 |
| | 0 | 2 | 25 | 150 | 570 |
Table 7: chlamydia concentration (IFU/ml)
| Time behind the bacterial infection (my god) | Contrast | The Radix Astragali | MOMP | The MOMP+ Radix Astragali | MOMP+ nanometer+ |
| 3 | 121678 | 130121 | 95461 | 70496 | 29530 |
| 9 | 85410 | 83456 | 81230 | 68950 | 20796 |
| 15 | 44565 | 47210 | 68950 | 47245 | 11298 |
| 21 | 30749 | 30918 | 20796 | 30489 | 9546 |
The foregoing description that this case provides not is that the present invention is carried out a kind of unique restriction, in actual applications, especially as carrier transfer bioactive substance treatment disease, or during as vaccine adjuvant, bioactive substance should comprise protein, or fat-soluble or soluble small molecular medicine, or polypeptide drug, described vaccine is meant protein or polypeptides matter, or antibody, or the gene genetic active matter.Their transfer mode should comprise mucosal drug delivery or by being administered systemically.Described mucosal drug delivery is meant the nasal mucosa administration, or vagina administration, or oral administration; Described being administered systemically is meant lumbar injection, or intravenous injection, or intramuscular injection.
Claims (3)
1, a kind of preparation method that contains the inorganic salt nano particles that bioactive substance and Chinese medicine ingredients seal is characterized in that:
A) preparation comprises the inorganic salt nano particles of bioactive substance:
After 0.01～1.0 mole saline solution, bioactive substance solution, 0.01～1.0 mole phosphoric acid solution mixed by 1: 1: 1, stirred 48 hours down at 4 ℃, supersound process 15 minutes, under 4 ℃, centrifugal 15 minutes of 8500rpm removes the supernatant that contains free bioactive substance solution, uses the deionized water suspended particles, concentration at 5mg/ml forms the inorganic salt nano particles that contains bioactive substance, and its size dimension is 100nm～800nm;
B) seal the active substance and the bioactive substance of Chinese medicine:
Under the room temperature, after will containing the active substance of inorganic salt particle, Chinese medicine of bioactive substance and bioactive substance and mixing by 1: 10: 10, concentration by 10mg/ml is suspended in pH value 7.0,0.01 in the phosphate buffer of mole, on shaking table, shook under 4 ℃ 2 hours, 4 ℃, centrifugal three times of 8500rpm, each 15 minutes, obtain the inorganic salt nano particles that contains bioactive substance and seal Chinese medicine ingredients;
Described saline solution is 0.01～1.0 mole a manganese chloride with the deionized water preparation, or magnesium chloride, or calcium chloride solution; Described bioactive substance is a protein; The active substance of described Chinese medicine is an astragalus polysaccharides, or Cordyceps polysaccharide, or pachyman, or lycium barbarum polysaccharide.
2, the preparation method that contains the inorganic salt nano particles that bioactive substance and Chinese medicine ingredients seal according to claim 1 is characterized in that: the inorganic salt nano particles that bioactive substance and Chinese medicine ingredients seal that contains of acquisition is stored in 4 ℃ or freeze-dried back.
3, the preparation method that contains the inorganic salt nano particles that bioactive substance and Chinese medicine ingredients seal according to claim 1, it is characterized in that: described protein is HSV-2 protein, or I (chlamydia) protein matter (MOMP), or human growth hormone (hGH).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100411635A CN100335038C (en) | 2005-07-25 | 2005-07-25 | Preparation process and application of inorganic salt nanometer particle containing biological active material and enveloped Chinese medicinal ingredient |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100411635A CN100335038C (en) | 2005-07-25 | 2005-07-25 | Preparation process and application of inorganic salt nanometer particle containing biological active material and enveloped Chinese medicinal ingredient |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1736366A CN1736366A (en) | 2006-02-22 |
| CN100335038C true CN100335038C (en) | 2007-09-05 |
Family
ID=36079439
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100411635A Expired - Fee Related CN100335038C (en) | 2005-07-25 | 2005-07-25 | Preparation process and application of inorganic salt nanometer particle containing biological active material and enveloped Chinese medicinal ingredient |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100335038C (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101148680B (en) * | 2007-10-23 | 2010-06-02 | 南京凯瑞尔纳米生物技术有限公司 | Method for introducing plasmid vector containing gene cure segment in cell by nano particles |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1635870A (en) * | 2000-10-09 | 2005-07-06 | 3M创新有限公司 | Medicinal aerosol formulations |
-
2005
- 2005-07-25 CN CNB2005100411635A patent/CN100335038C/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1635870A (en) * | 2000-10-09 | 2005-07-06 | 3M创新有限公司 | Medicinal aerosol formulations |
Non-Patent Citations (1)
| Title |
|---|
| 钙纳米粒子的制备及表征鉴定 郑金榆 等，南京医科大学学报（自然科学版），第24卷第4期 2003 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1736366A (en) | 2006-02-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP4233891A1 (en) | Targeting delivery system loaded with whole-cell components and use thereof | |
| WO2007113665A2 (en) | Polymerized solid lipid nanoparticles for oral or mucosal delivery of therapeutic proteins and peptides | |
| CN1840193A (en) | Nanometer capsule of anthracene nucleus anticancer antibiotic with polyethylene glycol-phospholipid | |
| CN1732918A (en) | Nanometer preparation of silybin and preparation method thereof | |
| CN103977407A (en) | Oral nanoparticle polypeptide composition tablets and preparation method thereof | |
| CN1582159A (en) | Preparing material of flavanol wood lipid with increased solubility | |
| WO2023040127A1 (en) | Use of cancer vaccine system based on whole cell components in preparation of drugs for cross-prevention or treatment of heterogeneous cancers | |
| CN100335038C (en) | Preparation process and application of inorganic salt nanometer particle containing biological active material and enveloped Chinese medicinal ingredient | |
| CN100435784C (en) | Method for preparing inorganic salt nanometer particles containing and packing traditional Chinese medicine and its application | |
| CN1775216A (en) | New formulation oxaliplatin liposome | |
| CN1823748A (en) | Medicinal preparation of coenzyme Q10 liposome and its preparation technology | |
| CN1264509C (en) | Precursor liposome preparation containing silybum marianum extract and its preparing process | |
| CN1698803A (en) | Compound notoginseng injection and its preparation process | |
| CN1301747C (en) | Protein medicine microcapsule and inhalational aerosol thereof | |
| CN1919339A (en) | Cucurbitacin nano preparation comprising protein, preparation method and use thereof | |
| CN107281163B (en) | Application of carboxyl compound in aspect of promoting oral absorption of drug-loaded nanoparticle microspheres | |
| CN1287857C (en) | Method for preparing specific nano vaccine for stomach cancer | |
| CN111514289A (en) | Synthesis method of novel coronavirus immune nanoparticle preparation | |
| AU2005283512A1 (en) | Permucosal composition and method of improving permucosal absorption | |
| CN1586488A (en) | Chitosan glycyrrhizic acid nano particle and its preparing method | |
| CN1504191A (en) | Cucurbitacin lipsome preparation method and formulation | |
| CN104436207B (en) | A kind of drug carrier system | |
| CN1465339A (en) | Nano-grade medicine microball and preparation process thereof | |
| CN103054805A (en) | Preparation method and application of injection for needle-free injection | |
| CN108210457B (en) | Medicinal application of collagen peptide, cucurbitacin oral micelle and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Owner name: NANJING JIANGKAI SCIENCE INDUSTRY CO., LTD.; HE Q Free format text: FORMER OWNER: JIANG QIAO; HE QING Effective date: 20071214 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20071214 Address after: Jiangsu Nanjing Nanjing hi tech Industrial Development Zone Software Park business villa 7, zip code: 210061 Co-patentee after: He Qing Patentee after: Nanjing Kai Kai Technology Industry Co., Ltd. Address before: Room 401, 11 Ma On Shan Road, Gulou District, Nanjing, Jiangsu, zip code: 210013 Co-patentee before: He Qing Patentee before: Jiangqiao |
|
| ASS | Succession or assignment of patent right |
Owner name: NANJING CARLE NANOMETER BIOLOGY TECHNOLOGY CO., L Free format text: FORMER OWNER: NANJING JIANGKAI SCIENCE INDUSTRY CO., LTD.; HE QING Effective date: 20071228 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20071228 Address after: Jiangsu Nanjing Nanjing hi tech Industrial Development Zone Software Park business villa 7, zip code: 210061 Patentee after: Kairuier Nana Biological Technology Co., Ltd., Nanjing Address before: Jiangsu Nanjing Nanjing hi tech Industrial Development Zone Software Park business villa 7, zip code: 210061 Co-patentee before: He Qing Patentee before: Nanjing Kai Kai Technology Industry Co., Ltd. |
|
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20070905 Termination date: 20150725 |
|
| EXPY | Termination of patent right or utility model |