Novel biological medicine based on porphyrin derivative and lipid and preparation process of novel biological medicine

Abstract

The invention discloses a novel biological medicine based on porphyrin derivatives and lipids and a preparation process of the novel biological medicine, and relates to the technical field of preparation of bioactive medicines. The medicine comprises protein containing natural porphyrin and derivatives thereof, and lipid containing natural or improved active ingredients, the substances are in a composition form of a mixture or a single substance and form liposomes or emulsions in a proper manner. According to the biological medicine, lipidosome or emulsion is used as a carrier, active ingredients such as porphyrin FS and hydrophobic febutadine a can be effectively transferred to a target position, the medicine transfer system can improve the stability and bioavailability of the medicine in a body, and the biological medicine is suitable for various administration routes such as oral cavity spraying, aerosol inhalation or solution inhalation and the like and has a wide application prospect. More flexible treatment selection is provided, the device is suitable for disease treatment needing local action, and the device is activated under irradiation of light with specific wavelength, so that the drug effect can be enhanced through control of an external light source.

Description

Technical Field

[0001] The present invention belongs to the technical field of preparation of bioactive drugs, and in particular relates to a novel biopharmaceutical based on porphyrin derivatives and lipids and a manufacturing process thereof. Background Art

[0002] Porphyrin is a macrocyclic compound containing four pyrrole molecules. Due to its unique structure and special properties, it has been widely used in biochemistry, medicine, analytical chemistry, synthetic chemistry, materials science and other fields in recent years.

[0003] Current research on different types of porphyrins includes modifying protoporphyrin with polyethylene glycol to prepare water-soluble porphyrin derivatives, and using nitrobenzaldehyde and pyrrole as raw materials to prepare tetraaminophenylporphyrin. Furthermore, porphyrin-containing siloxanes have also been prepared, but these methods simply dope the porphyrin into the siloxane, where the porphyrin acts as a thermal stabilizer. Different porphyrins or varying amounts of porphyrin are added as needed to improve the thermal stability of the siloxane. However, these methods do not effectively improve the processing properties of porphyrins.

[0004] To this end, we provide new biopharmaceuticals based on porphyrin derivatives and lipids and their manufacturing processes to solve the above problems. Summary of the Invention

[0005] To solve the above technical problems, the present invention is achieved through the following technical solutions:

[0006] The present invention is a novel biopharmaceutical based on porphyrin derivatives and lipids, which comprises a protein containing natural porphyrin and its derivatives, and a lipid with a natural or modified active ingredient;

[0007] The above substances are in the form of mixtures or individual substances and are formed into liposomes or emulsions by appropriate means.

[0008] The present invention is further configured such that the emulsion contains:

[0009] 0.001% to 1% porphyrin substance FS;

[0010] 0.001% to 1% hydrophobic feoofitin a;

[0011] 0.01% to 10% lipid as an emulsifier;

[0012] and acceptable auxiliary ingredients, taste and aroma additives, and water.

[0013] The present invention is further configured such that the lipid of the active ingredient is derived from cyanobacteria of the genus Spirulina, plants of the genus Urea or phospholipids derived from cholesterol.

[0014] The present invention is further provided that the drug is prepared in liquid form, has a higher molar concentration of lipid content, is suitable for oral spraying, aerosol inhalation or solution inhalation, and can be activated under irradiation of light of a specific wavelength.

[0015] The present invention is further configured such that the drug also contains lecithin and cholesterol, with the ratio of the two being 7:3.

[0016] The manufacturing process of the novel biopharmaceutical based on porphyrin derivatives and lipids includes the following steps:

[0017] preparing proteins and their derivatives containing natural porphyrins;

[0018] Preparation of active lipids of natural or modified origin;

[0019] The above ingredients are combined to form liposomes or emulsions.

[0020] The present invention is further configured such that the extraction process of the porphyrin substance FS comprises the following steps:

[0021] Extraction of raw porphyrin;

[0022] acid treatment to remove magnesium ions;

[0023] Washing with organic solvents (e.g., hexane, acetone), hydrochloric acid, and citric acid buffers;

[0024] Phosphorylation was performed in vacuo;

[0025] Combined with differential chromatography to improve extraction efficiency.

[0026] The present invention is further configured to mix the porphyrin solution with ethanol, purified water and glycerol (20%) by high-intensity mixing in a turbulent flow mode, thereby obtaining an emulsion with a specific ratio;

[0027] The specific steps include mixing the alcohol solution of FS with a hydro-alcoholic mixture and glycerol or Tyzol (surfactant), followed by further mixing under turbulent conditions.

[0028] The present invention is further configured such that the types of liposomes in the emulsion include multilamellar vesicles (MLL), large unilamellar vesicles (BOL) and small unilamellar vesicles;

[0029] Hairy liposomes can be formed by using phospholipids; other lipids such as phosphatidyldiethanolamine or phosphatidyldianiline can also be used to prepare hairy liposomes.

[0030] The present invention has the following beneficial effects:

[0031] 1. The biopharmaceuticals containing porphyrin derivatives and lipids provided by the present invention utilize liposomes or emulsions as carriers to effectively deliver active ingredients (such as porphyrin substance FS and hydrophobic pheophytin a) to the target site. This drug delivery system can improve the stability and bioavailability of the drug in the body.

[0032] 2. The biopharmaceuticals provided by the present invention, which contain porphyrin derivatives and lipids, are prepared in liquid form and are suitable for a variety of administration routes, including oral spray, aerosol inhalation, or solution inhalation, providing more flexible treatment options. They are particularly suitable for the treatment of diseases that require local effects and are activated under irradiation with light of a specific wavelength. Thus, the drug effect can be enhanced by controlling an external light source, providing a non-invasive treatment method.

[0033] 3. The active ingredients in the biopharmaceuticals containing porphyrin derivatives and lipids provided by the present invention are derived from phospholipids derived from cyanobacteria, plants, or cholesterol in nature. These natural ingredients generally have good biocompatibility and low toxicity risk, which increases the safety of drug use. In addition, by using different lipid materials and technologies (such as phosphatidylserine proteins), various types of liposomes such as multi-hairy liposomes, large unilamellar liposomes, and small unilamellar liposomes can be prepared, providing the possibility of optimizing the release characteristics and targeting ability of drugs.

[0034] Of course, any product implementing the present invention does not necessarily need to achieve all of the advantages described above at the same time. DETAILED DESCRIPTION

[0035] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0036] Example

[0037] The present invention is a novel biopharmaceutical based on porphyrin derivatives and lipids, which comprises a protein containing natural porphyrin and its derivatives, and a lipid with a natural or modified active ingredient;

[0038] The above substances are in the form of a mixture or a single substance, and are formed into liposomes or emulsions by appropriate means;

[0039] The emulsion contains:

[0040] 0.001% to 1% porphyrin substance FS;

[0041] 0.001% to 1% hydrophobic feoofitin a;

[0042] 0.01% to 10% lipid as an emulsifier;

[0043] and acceptable auxiliary ingredients, taste and aroma additives, and water.

[0044] Specifically, the drug also contains lecithin and cholesterol in a ratio of 7:3; in addition, the lipids of the active ingredients of the drug are derived from cyanobacteria of the genus Spirulina, uric acid plants or phospholipids derived from cholesterol.

[0045] The drug is prepared in liquid form with a higher molar concentration of lipids, suitable for oral spray, aerosol inhalation or solution inhalation, and can be activated by exposure to light of a specific wavelength.

[0046] The manufacturing process of the novel biopharmaceutical based on porphyrin derivatives and lipids includes the following steps:

[0047] preparing proteins and their derivatives containing natural porphyrins;

[0048] Preparation of active lipids of natural or modified origin;

[0049] The above ingredients are combined and mixed with high intensity in turbulent mode to mix the porphyrin solution with ethanol, purified water and glycerol (20%) to obtain an emulsion with a specific ratio; the specific steps include mixing the alcohol solution of FS with a water-fat-alcohol mixture and glycerol or Tyzol (surfactant), followed by further mixing under turbulent conditions.

[0050] The content of the emulsion is FS of porphyrin substance, content of hydrophobic feoofitin a is 0.001-1%, content of lipid as emulsifier is 0.01-10%, and acceptable auxiliary ingredients, taste and aroma additives and water, so that these components form Mll drug or related emulsion, the lipid of active ingredient can be obtained from cyanobacteria of Spirulina or uric acid plant or phospholipid composed of cholesterol, the Mll drug or related emulsion is made into a liquid with a higher molar concentration of lipid content for use; the types of liposomes in the emulsion include multilamellar liposomes (MLL), large unilamellar liposomes (BOL) and small unilamellar liposomes; multilamellar liposomes can be formed by using phospholipidamide protein; other lipids such as phosphatidyldiethanolamine or phosphatidyldianiline can also be used to prepare multilamellar liposomes.

[0051] The extraction process of porphyrin substance FS includes the following steps:

[0052] Extraction of raw porphyrin;

[0053] acid treatment to remove magnesium ions;

[0054] Washing with organic solvents (e.g., hexane, acetone), hydrochloric acid, and citric acid buffers;

[0055] Phosphorylation was performed in vacuo;

[0056] Combined with differential chromatography to improve extraction efficiency.

[0057] Examples of applications of biopharmaceuticals containing porphyrin derivatives and lipids are as follows:

[0058] Example 1: Steps for extracting Theophiphin from the plant Urtica dioica

[0059] 1) Dried nettles are ground into powder and sifted through a sieve;

[0060] 2) Add 6.25 liters of sodium dihydrogen phosphate aqueous solution per 1.25 kg of crushed nettles and stir and dissolve the mixture at room temperature for 2 hours;

[0061] 3) Freeze the resulting mixture for 12 hours and then centrifuge it at 0°C for 10 minutes to remove water;

[0062] 4) Treat the sediment with acetone (3 times, 2.5 L each time) to completely extract the mixture of chlorophylls A and B;

[0063] 5) Collect the liquid extract after centrifugation, filter to remove mechanical impurities, and measure the final extract volume.

[0064] As for the treatment of chlorophylls A and B, 20 mL of concentrated hydrochloric acid was first added to the extract. After stirring at room temperature for 20 minutes, the pH was adjusted to 6-8 using 20% ​​sodium hydroxide solution. After confirming the pH value with universal indicator paper, the desired compounds were isolated by adsorption on cotton or wool, and subsequent processing steps such as rotary evaporation were performed.

[0065] Example 2: Preparation of Alcoholic Lecithin Solution

[0066] The porphyrin solution, ethanol, purified water, and glycerol (20%) were mixed under high intensity in a turbulent flow mode to obtain an emulsion containing 0.001% porphyrin and 0.01% lecithin;

[0067] Lecithin and cholesterol were prepared in a ratio of 7:3 and mixed under the same conditions to obtain an emulsion containing 0.01% porphyrin and 0.1% lipid.

[0068] Example 3: Determining Liposome Parameters

[0069] The emulsion sample was applied to a Sephadex G-50 column and eluted with physiological solution. Fractions at two different absorption wavelengths were collected for analysis of the concentration of FS in the liposomes and its inclusion degree.

[0070] Example 4: Application of Emulsion

[0071] The emulsion is suitable for administration to the oral mucosa or intranasal cavity (0.04 ml in each nostril). After administration, it can be irradiated with a specific light source, such as a mercury lamp or LED light source, to enhance the drug effect.

[0072] Throughout this specification, references to terms such as "one embodiment," "example," or "specific example" indicate that the specific features, structures, materials, or characteristics described in conjunction with that embodiment or example are included in at least one embodiment or example of the present invention. In this specification, schematic representations of these terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

[0073] The preferred embodiments of the present invention disclosed above are intended only to help illustrate the present invention. These preferred embodiments do not exhaustively describe all details, nor do they limit the present invention to the specific embodiments described. Obviously, many modifications and variations are possible based on the content of this specification. These embodiments are selected and described in detail in this specification to better explain the principles and practical applications of the present invention, thereby enabling those skilled in the art to better understand and utilize the present invention. The present invention is limited only by the claims and their full scope and equivalents.

Claims

1. A novel biopharmaceutical based on porphyrin derivatives and lipids, characterized in that: The drug comprises proteins containing natural porphyrins and their derivatives, as well as lipids of natural or modified active ingredients; The above substances are in the form of mixtures or individual substances and are formed into liposomes or emulsions by appropriate means.

2. The novel biopharmaceutical based on porphyrin derivatives and lipids according to claim 1, characterized in that: The emulsion contains: 0.001% to 1% porphyrin substance FS; 0.001% to 1% hydrophobic feoofitin a; 0.01% to 10% lipid as an emulsifier; and acceptable auxiliary ingredients, taste and aroma additives, and water.

3. The novel biopharmaceutical based on porphyrin derivatives and lipids according to claim 1, characterized in that: The lipid of the active ingredient is derived from cyanobacteria of the genus Spirulina, phospholipids derived from plants of the genus Urea or cholesterol.

4. The novel biopharmaceutical based on porphyrin derivatives and lipids according to claim 1, characterized in that: The drug is prepared in liquid form with a higher molar concentration of lipids, suitable for oral spray, aerosol inhalation or solution inhalation, and can be activated by exposure to light of a specific wavelength.

5. The novel biopharmaceutical based on porphyrin derivatives and lipids according to claim 1, characterized in that: The drug also contains lecithin and cholesterol in a ratio of 7:

3.

6. The manufacturing process of the novel biopharmaceutical based on porphyrin derivatives and lipids according to any one of claims 1 to 5, characterized in that: The following steps are involved: preparing proteins and their derivatives containing natural porphyrins; Preparation of active lipids of natural or modified origin; The above ingredients are combined to form liposomes or emulsions.

7. The manufacturing process of the novel biopharmaceutical based on porphyrin derivatives and lipids according to claim 6, characterized in that: The extraction process of porphyrin substance FS includes the following steps: Extraction of raw porphyrin; acid treatment to remove magnesium ions; Washing with organic solvents (e.g., hexane, acetone), hydrochloric acid, and citric acid buffers; Phosphorylation was performed in vacuo; Combined with differential chromatography to improve extraction efficiency.

8. The manufacturing process of the novel biopharmaceutical based on porphyrin derivatives and lipids according to claim 6, characterized in that: The porphyrin solution was mixed with ethanol, purified water, and glycerol (20%) under high-intensity mixing in turbulent mode to obtain an emulsion with a specific ratio; The specific steps include mixing the alcohol solution of FS with a hydro-alcoholic mixture and glycerol or Tyzol (surfactant), followed by further mixing under turbulent conditions.

9. The manufacturing process of the novel biopharmaceutical based on porphyrin derivatives and lipids according to claim 6, characterized in that: The types of liposomes in the emulsions included multilamellar vesicles (MLLs), large unilamellar vesicles (BOLs), and small unilamellar vesicles; Hairy liposomes can be formed by using phospholipids; other lipids such as phosphatidyldiethanolamine or phosphatidyldianiline can also be used to prepare hairy liposomes.