CN112006990A - Nano-targeting transdermal method - Google Patents
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- CN112006990A CN112006990A CN202010689527.5A CN202010689527A CN112006990A CN 112006990 A CN112006990 A CN 112006990A CN 202010689527 A CN202010689527 A CN 202010689527A CN 112006990 A CN112006990 A CN 112006990A
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- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
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Abstract
The invention provides a nano-targeting transdermal method, which comprises the following steps: lecithin, soluble protein, targeting polypeptide and soluble metal salt; the method comprises the following steps: the lecithin is dissolved by an organic solvent to prepare a lecithin solution, and the soluble protein, the targeting polypeptide and the soluble metal salt are respectively dissolved by water to prepare a protein solution, a polypeptide solution and a metal salt solution; adding the protein solution and the polypeptide solution into the lecithin solution, and performing vortex oscillation and ultrasonic treatment to obtain a solution A; adding the metal salt solution into the solution A, and performing vortex oscillation and ultrasonic treatment to obtain a solution B; the solution B is sealed, stirred in a water bath, heated, oscillated in a vortex mode and subjected to ultrasonic wave, pure water is added, and organic solvent is removed through rotary evaporation to obtain a transparent nano targeting ligand solution; the transparent nano targeting ligand solution is frozen and dried; the invention has the advantages of simple and easily obtained raw materials, simple preparation process, low cost and wide applicability, and can realize large-scale industrial production.
Description
Technical Field
The invention relates to the technical field of biomedicine, in particular to a nano-targeting transdermal method.
Background
Cancer is currently the most common and serious disease seriously threatening human health and life, and the current drug therapy for cancer still faces a plurality of bottleneck problems, such as high recurrence rate, rapid drug resistance, high toxic and side effects, low drug effect and the like. The existing anticancer drugs used in clinic have the following problems: the sensitization medicine has high toxicity and poor targeting property, or the inconvenience of clinical application is increased by matching daily radiotherapy with daily medication, and the like. The anti-cancer medicine taking cisplatin as an example has a good anti-cancer effect, can be used as a first-line medicine for chemotherapy, and meanwhile, the cisplatin is a high-efficiency radiotherapy sensitizer and can remarkably enhance the radiotherapy effect. However, cisplatin causes great damage to normal cells of a human body while killing tumor cells, toxic reactions mainly include nephrotoxicity, digestive tract toxicity, bone marrow suppression, neurotoxicity, ototoxicity and the like, and serious toxic and side effects cause that patients cannot continue drug therapy.
To address this problem, researchers have envisioned and achieved some success by targeting anticancer drugs directly to the tumor site through formulation means, such as liposomes. The liposome technology is a fourth generation targeted drug delivery technology which is similar to biological missile, and by utilizing the unique characteristics of liposome, the drugs with large side effect, poor stability in blood and rapid degradation are wrapped in the liposome, and according to the larger vascular endothelial cell gap of the focus part of a human body, the liposome drugs can reach the focus part through the gap, so that the targeted release drug delivery is realized, and the clinical treatment is safe and effective. The liposome as the antitumor drug carrier has the characteristics of changing the distribution of the drug in tissues, improving the therapeutic index and the bioavailability, increasing the curative effect and reducing the toxic and side effects. The long-acting targeted liposome is used as a drug carrier, so that the diffusion of the liposome to a tumor area is improved, the amount of drugs reaching the tumor is increased, and the cure rate is improved. Liposomes have been used clinically as carriers of drugs for many years, such as doxorubicin liposomes. However, the biggest defects of the liposome are chemical and physical instability, the liposome vesicle is easy to break, the anti-cancer drug is leaked out early before reaching the tumor site, the slow release effect is not achieved, and the wide application of the liposome in clinic is seriously influenced. Just like the car in winter north has installed the one deck tyre chain additional, when increasing car stability, has also increased the rigidity of tire. This increases the difficulty of endocytosis of the silica by the tumor cells. As a drug carrier, the hydrolysis by-product of ethanol increases the side effects of the silica or increases the separation step. Unlike the siliceous body, the biomembrane ligand is prepared with human and animal cell membrane as material and through forming coordination bond between iron and other metal ion and amino or phosphate group in protein and phospholipid, forming inside liposome numerous organic-metal coordination bonds to stabilize liposome, and adding elastic fiber network structure inside automobile wheel. Particularly, the iron complex is easy to metabolize in a human body, so that the biocompatibility is better, and the cytotoxicity is lower. The biomembrane gametocyte is delivered to the tumor tissue under the mediation of the targeting polypeptide and is combined with a tumor cell receptor. The gametocyte has the function of specifically targeting target tumor tissues or cells, and can play four anticancer functions: (1) can be used as a carrier of lipophilic and hydrophilic chemotherapeutic drugs and anticancer genes; (2) under the external excitation of light, sound, wave and the like, the medicine can simultaneously generate heat with the effect of killing tumors and active oxygen-drug-free photothermal therapy and photodynamic therapy; (3) the magnetic resonance imaging system has strong magnetic resonance image enhancement and photoacoustic imaging functions, and can be used for diagnosis of early tumors and intraoperative navigation; (4) can be used as sensitizer in tumor thermotherapy.
Disclosure of Invention
In this summary, concepts in a simplified form are introduced that are further described in the detailed description section. This summary of the invention is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used as an aid in determining the scope of the claimed subject matter.
In order to at least partially solve the technical problems, the invention provides a nano-targeting transdermal method, which comprises the following raw materials in parts by weight:
160-250 parts of lecithin, 40-60 parts of soluble protein, 10-50 parts of targeting polypeptide and 5-20 parts of soluble metal salt;
the nano-targeting transdermal method comprises the following steps:
(1) the lecithin is dissolved by an organic solvent to prepare a lecithin solution, and the soluble protein, the targeting polypeptide and the soluble metal salt are respectively dissolved by water to prepare a protein solution, a polypeptide solution and a metal salt solution;
(2) adding the protein solution and the polypeptide solution into the lecithin solution, performing vortex oscillation for 5-20 min, and performing ultrasonic treatment for 10-30 min to obtain a solution A;
(3) adding a metal salt solution into the solution A, performing vortex oscillation for 5-20 min, and performing ultrasonic treatment for 10-30 min to obtain a solution B;
(4) sealing the solution B, stirring and heating in a water bath at 30-55 ℃ for 2-10 h, carrying out vortex oscillation for 5-10 min, carrying out ultrasonic wave for 10-30 min, adding 10-20 parts of ultrapure water, and carrying out rotary evaporation to remove the organic solvent to obtain a transparent nano targeting ligand solution;
(5) and (3) freeze-drying the transparent nano targeting ligand solution.
Further, the targeted polypeptide is a polypeptide formed by dehydration polymerization of 5-31 amino acids.
Further, the soluble proteins include: one or more of soluble protein extracted from plant tissue, soluble protein extracted from animal, human serum albumin, and artificial protein synthesized by recombinant gene.
Further, the soluble metal salt is one or a combination of more of ferric chloride hexahydrate, anhydrous ferric chloride, ferrous sulfate, ammonium ferrous sulfate, ferrous chloride, potassium ferrocyanide, potassium ferricyanide, copper chloride, copper sulfate, manganese acetate, manganese sulfate, gadolinium trichloride, ferric manganese chlorophyllin, zinc sulfate, nickel sulfate, bismuth trichloride, transition metal salt and rare earth metal salt.
Further, the transition metal salt solution is lanthanum nitrate, cerium nitrate, praseodymium nitrate, neodymium nitrate, promethium nitrate, samarium nitrate, europium nitrate, gadolinium nitrate, terbium nitrate, dysprosium nitrate, holmium nitrate, erbium nitrate, thulium nitrate, ytterbium nitrate, lutetium nitrate, scandium nitrate, and yttrium nitrate.
Compared with the prior art, the invention has the technical effects that: (1) the preparation body of the invention has the capability of easily penetrating biological membranes and other physiological tissue barriers, particularly has a plurality of active groups on the surface, such as amino, carboxyl, hydroxyl and the like, and is easily coupled with tumor specific targeting polypeptide, cell penetrating peptide and other small molecules, so that the preparation body has a synergistic targeting effect. In addition, the particle size of the ligand body is 50-150 nm, the number of particles formed by small particle size and the number of atoms on the surface layer of the formed nano particles are greatly increased, the covering capability and the attachment capability are enhanced, and the contact area between the particles and the skin is greatly increased; (2) the biomembrane nano targeting ligand can be used as a carrier of lipophilic and hydrophilic chemotherapeutic drugs and anticancer genes, and improves the biocompatibility, targeting property, degradability, bioavailability, stability and the like of the drugs; (3) the invention also has the advantages of simple and easily obtained raw materials, simple preparation process, low cost and wide applicability, and can be used for large-scale industrial production.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications.
Example 1A Nanogated transdermal method
Raw materials: 160 parts of lecithin, 40 parts of bovine serum albumin, 10 parts of targeting polypeptide and 5 parts of ferrous sulfate;
the method comprises the following steps:
(1) the lecithin is dissolved by chloroform to prepare a lecithin chloroform solution, and the bovine serum albumin, the targeting polypeptide and the ferrous sulfate are respectively dissolved by water to obtain a protein solution, a polypeptide solution and a ferric sulfate solution;
(2) adding the polypeptide solution and the protein solution into a lecithin chloroform solution, performing vortex oscillation for 20min at the rotation speed of 1000r/min, and performing ultrasonic treatment at 40kHz for 10min to obtain a suspension A;
(3) adding ferrous sulfate solution into the solution A, performing vortex oscillation for 20min at the rotation speed of 1000r/min, and performing ultrasonic treatment at 80kHz for 20min to obtain suspension B;
(4) placing the suspension B in a water bath at 40 ℃, stirring for 2.5h, adding 10 parts of water, performing vortex oscillation at the rotation speed of 1000r/min for 10min, performing ultrasonic treatment at the frequency of 40kHZ for 20min, fully mixing uniformly, and performing rotary evaporation to remove the organic solvent to obtain a transparent nano targeting ligand solution;
(5) the transparent nano targeting ligand solution is frozen and dried into powder at the temperature of minus 50 ℃.
Example 2A Nano-targeting transdermal method
Raw materials: 200 parts of phospholipid, 50 parts of human serum albumin, 30 parts of targeting polypeptide, 5 parts of manganese sulfate and 10 parts of potassium ferrocyanide;
the method comprises the following steps:
(1) dissolving lecithin in chloroform-ethanol (volume ratio of 2: 1) to obtain lecithin chloroform-ethanol solution, and dissolving human serum albumin, target polypeptide, manganese sulfate and ferrous sulfate in water to obtain protein solution, polypeptide solution, manganese sulfate solution and ferrous sulfate solution;
(2) adding the polypeptide solution and the protein solution into a chloroform-ethanol solution of the phospholipid, performing vortex oscillation for 10min at a rotation speed of 1500r/min, and performing ultrasonic treatment at 80kHz for 20min to obtain a suspension A;
(3) adding the manganese sulfate solution and the sulfuric acid sub-solution into the solution A, performing vortex oscillation for 10min at the rotating speed of 1500r/min, and performing ultrasonic treatment at 80kHz for 20min to obtain a suspension B;
(4) placing the suspension B in a water bath at 50 ℃, stirring for 3h, adding 10 parts of water, performing vortex oscillation at the rotation speed of 1500r/min for 8min, performing ultrasonic treatment at the frequency of 80kHZ for 20min, fully mixing uniformly, and performing rotary evaporation to remove the organic solvent to obtain a transparent nano targeting ligand solution;
(5) the transparent nano targeting ligand solution is frozen and dried into powder at the temperature of minus 60 ℃.
Example 3A Nano-targeting transdermal method
Raw materials: 250 parts of lecithin, 60 parts of soybean albumin, 50 parts of targeting polypeptide, 3 parts of potassium ferrocyanide, 4 parts of ferric trichloride salt and 1 part of manganese sulfate;
the method comprises the following steps:
(1) dissolving the lecithin with cyclohexane to prepare a lecithin cyclohexane solution, and dissolving the soybean albumin, the target polypeptide, the potassium ferrocyanide, the ferric trichloride salt and the manganese sulfate with water respectively to obtain a protein solution, a polypeptide solution, the potassium ferrocyanide, the ferric trichloride solution and a manganese sulfate solution;
(2) adding the polypeptide solution and the protein solution into the lecithin cyclohexane solution, performing vortex oscillation for 5min at the rotating speed of 3000r/min, and performing ultrasonic treatment at 100kHz for 20min to obtain a suspension A;
(3) adding the manganese sulfate solution and the sulfuric acid sub-solution into the solution A, performing vortex oscillation for 5min at the rotating speed of 3000r/min, and performing 100kHz ultrasonic oscillation for 20min to obtain a suspension B;
(4) placing the suspension B in a water bath at 35 ℃, stirring for 10h, adding 15 parts of water, performing vortex oscillation at the rotation speed of 3000r/min for 10min, performing ultrasonic treatment at the frequency of 100kHZ for 20min, fully mixing uniformly, and performing rotary evaporation to remove the organic solvent to obtain a transparent nano targeting ligand solution;
(5) the transparent nano targeting ligand solution is frozen and dried into powder at the temperature of-70 ℃.
Specifically, the targeted polypeptide is a polypeptide prepared by dehydrating and polymerizing 5-31 amino acids.
In particular, the soluble proteins include: one or more of soluble protein extracted from plant tissue, soluble protein extracted from animal, human serum albumin, and artificial protein synthesized by recombinant gene.
Specifically, the soluble metal salt is one or a combination of more of ferric trichloride hexahydrate, anhydrous ferric trichloride, ferrous sulfate, ammonium ferrous sulfate, ferrous chloride, potassium ferrocyanide, potassium ferricyanide, copper chloride, copper sulfate, manganese acetate, manganese sulfate, gadolinium trichloride, manganese ferrite chlorophyllin, zinc sulfate, nickel sulfate, bismuth trichloride, transition metal salt and rare earth metal salt.
Specifically, the transition metal salt solution is lanthanum nitrate, cerium nitrate, praseodymium nitrate, neodymium nitrate, promethium nitrate, samarium nitrate, europium nitrate, gadolinium nitrate, terbium nitrate, dysprosium nitrate, holmium nitrate, erbium nitrate, thulium nitrate, ytterbium nitrate, lutetium nitrate, scandium nitrate, and yttrium nitrate.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (5)
1. A nano-targeting transdermal method is characterized in that the raw materials are as follows according to parts by weight:
160-250 parts of lecithin, 40-60 parts of soluble protein, 10-50 parts of targeting polypeptide and 5-20 parts of soluble metal salt.
2. The nano-targeted transdermal method according to claim 1, wherein the preparation process comprises:
(1) the lecithin is dissolved by an organic solvent to prepare a lecithin solution, and the soluble protein, the targeting polypeptide and the soluble metal salt are respectively dissolved by water to prepare a protein solution, a polypeptide solution and a metal salt solution;
(2) adding the protein solution and the polypeptide solution into the lecithin solution, and performing vortex oscillation and ultrasonic treatment to obtain a solution A;
(3) adding the metal salt solution into the solution A, and performing vortex oscillation and ultrasonic treatment to obtain a solution B;
(4) the solution B is sealed, stirred in a water bath, heated, oscillated in a vortex mode and subjected to ultrasonic wave, pure water is added, and organic solvent is removed through rotary evaporation to obtain a transparent nano targeting ligand solution;
(5) and (3) freeze-drying the transparent nano targeting ligand solution.
3. The nano-targeting transdermal method according to claim 1, wherein the targeting polypeptide is a polypeptide obtained by dehydration polymerization of 5 to 31 amino acids.
4. The nano-targeted transdermal method according to claim 1, wherein the soluble protein comprises: one or more of soluble protein extracted from plant tissue, soluble protein extracted from animal, human serum albumin, and artificial protein synthesized from recombinant gene.
5. The nano-targeted transdermal method according to claim 1, wherein the soluble metal salt is one or more of ferric chloride hexahydrate, ferric chloride anhydrous, ferrous sulfate, ammonium ferrous sulfate, ferrous chloride, potassium ferrocyanide, potassium ferricyanide, copper chloride, copper sulfate, manganese acetate, manganese sulfate, gadolinium trichloride, ferric manganese chlorophyllin salt, zinc sulfate, nickel sulfate, bismuth trichloride, transition metal salts, and rare earth metal salts.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112708415A (en) * | 2020-12-23 | 2021-04-27 | 国家纳米科学中心 | Iron-containing nano probe, preparation method and synchronous radiation application |
| CN116590234A (en) * | 2023-07-12 | 2023-08-15 | 中国农业大学 | Method for promoting endocytosis of plant albumin and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107335063A (en) * | 2017-05-27 | 2017-11-10 | 广西玉林魔树树棉科技有限公司 | A kind of Anti-cancer biologic film nano target is with plastid and preparation method and application |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107335063A (en) * | 2017-05-27 | 2017-11-10 | 广西玉林魔树树棉科技有限公司 | A kind of Anti-cancer biologic film nano target is with plastid and preparation method and application |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112708415A (en) * | 2020-12-23 | 2021-04-27 | 国家纳米科学中心 | Iron-containing nano probe, preparation method and synchronous radiation application |
| CN116590234A (en) * | 2023-07-12 | 2023-08-15 | 中国农业大学 | Method for promoting endocytosis of plant albumin and application thereof |
| CN116590234B (en) * | 2023-07-12 | 2023-09-29 | 中国农业大学 | Method for promoting endocytosis of plant albumin and application thereof |
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