CN112891556A - Oral nanogel of monoclonal antibody medicines and preparation method thereof - Google Patents
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- A61K47/00—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
- 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
- A61K47/56—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 an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/61—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 an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule the organic macromolecular compound being a polysaccharide or a derivative thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
- A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—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
- 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
- A61K47/54—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 an organic compound
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—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
- 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
- A61K47/69—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 conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6903—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 conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being semi-solid, e.g. an ointment, a gel, a hydrogel or a solidifying gel
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
Abstract
The invention discloses a monoclonal antibody medicine oral nano gel preparation and a preparation method thereof. The monoclonal antibody drug oral nano gel is prepared by crosslinking a monoclonal antibody drug by a disulfide bond crosslinking agent (2- [ (2- { [ (4-nitrophenoxy) carbonyl ] oxy } ethyl) disulfide group ] ethyl 4-nitrobenzoate) or a homolog thereof to form a nano cross-linked substance, and then connecting hyaluronic acid to the surface of the nano cross-linked substance through chemical synthesis. The monoclonal antibody medicine oral nano gel can directly transport the antibody medicine to an inflammation part by adopting an oral administration means, can avoid adverse reaction caused by systemic administration of the medicine, and simultaneously increases the compliance of patients. The nanogel prepared by the method can be gathered at the inflammatory part of the intestinal tract through a targeting effect, and simultaneously, the S-S bond crosslinked by the antibody can be degraded in a responsive manner under the intestinal tract environment to release the drug, so that the targeted and positioned release of the drug at the inflammatory part of the intestinal tract is realized.
Description
(I) technical field
The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a monoclonal antibody drug oral nanogel and a preparation method thereof.
(II) background of the invention
Inflammatory Bowel Disease (IBD) is a chronic nonspecific inflammatory disease of intestinal tract with unknown etiology, mainly comprises crohn's disease and ulcerative colitis, which affect the intestinal mucosa, cause inflammation, erosion, ulcer and heavy bleeding of intestinal tract, and seriously affect the life quality of patients. Clinically, anti-TNF-alpha monoclonal antibody (such as infliximab, adalimumab, certolizumab ozogamicin and the like) is tried to treat IBD, and good curative effect is achieved. However, anti-TNF- α mabs are usually administered by injection, and this systemic administration results in systemic inhibition of TNF- α in the absence of selective action of the drug at the target site, resulting in a series of adverse effects. If the antibody drug can be directly transported to an inflammation part to act with TNF-alpha by means of oral administration, adverse reaction caused by systemic administration of the drug can be avoided.
However, the main problems faced by oral protein pharmaceutical preparations are that they are easily degraded by the acid-base environment in the digestive tract and various proteolytic enzymes to lose biological activity; secondly, the medicine has larger molecular weight and small fat solubility, and is difficult to be absorbed through the biological mucous membrane of the gastrointestinal tract. It is noted that anti-TNF- α mabs can directly bind to TNF- α at the site of IBD lesions without being absorbed through gastrointestinal biofilms into the blood circulation, i.e. exert a pharmacodynamic effect, which provides the possibility for oral administration of such mabs. In addition, the permeability of intestinal mucosa of IBD lesion sites is enhanced, which is beneficial for the penetration of a nano-carrier system into the mucosa of an inflammation area. For a long time, pharmaceutical workers at home and abroad have adopted various strategies to improve the stability of oral protein drugs, such as the application of enzyme inhibitors, the development of novel submicron carriers and chemical structure modification. Prolonged use of enzyme inhibitors can cause irreversible mucosal damage and gastrointestinal digestive disorders. Chemical structure modifications affect the activity of the drug and its in vivo metabolic kinetic properties. In contrast, the nano oral drug delivery system has unique advantages in protecting the stability of protein drugs in the gastrointestinal tract.
For protein drug nano-preparations, how to improve the entrapment capacity of water-soluble biomacromolecule drugs is always a difficulty in designing nano-drug carrier systems. Protein drugs are mainly encapsulated through electrostatic adsorption, coating, embedding and other actions between the protein drugs and a carrier, and the problems of low drug loading rate and low encapsulation rate exist mostly. In addition, some existing preparation methods also have certain influence on the activity of the protein drug, such as the use of an organic solvent in an emulsion solvent volatilization method.
How to realize the targeted and positioned release of the protein drug in the intestinal tract is also the key point of the design of the nanogel drug carrier system. For inflammatory bowel diseases, inflammatory sites in the intestinal tract are segmental, small in area and dispersed, and certain difficulty is brought to targeted and positioned release of drugs.
At present, no oral nano-administration preparation of the anti-TNF-alpha monoclonal antibody is seen in the market, and no research report of similar pharmaceutical preparations is seen, so that the development of the medicine has important clinical significance and wide market prospect.
Disclosure of the invention
Based on the defects in the prior art and the market demand, the invention provides a monoclonal antibody, in particular to an anti-TNF-alpha monoclonal antibody drug oral nanogel and a preparation method thereof.
In order to solve the technical problems, the invention adopts the technical scheme that:
an oral nano-gel preparation of monoclonal antibody medicines is obtained by the following method: and (2- { [ (4-nitrophenoxy) carbonyl ] oxy } ethyl) disulfide group ] ethyl 4-nitrobenzoate (hereinafter referred to as cross-linking agent) or (2- [ (2- { [ (4-nitrophenoxy) carbonyl ] oxy } propyl) disulfide group ] ethyl 4-nitrobenzoate) and the like, cross-linking the monocistronic drug to form a nano cross-linked substance, and connecting hyaluronic acid to the surface of the nano cross-linked substance through chemical synthesis to obtain the monoclonal drug oral nanogel.
According to the invention, through the cross-linking agent with the intestinal responsiveness breaking capacity, the monoclonal antibody drug is cross-linked to form a nano cross-linked substance with excellent stability, and then hyaluronic acid is modified to the surface of the nano cross-linked substance to form nano gel with negative points, and the nano gel can be gathered to an inflammation part, so that the monoclonal antibody drug oral nano gel preparation with high drug loading capacity and encapsulation efficiency and inflammation targeting performance is finally obtained.
Preferably, the monoclonal antibody drug is an anti-TNF-alpha monoclonal antibody drug.
Preferably, the hyaluronic acid is aminated hyaluronic acid with a molecular weight range of 10-100 kDa, and the ratio of the amount of hyaluronic acid to the amount of monoclonal antibody substance is 1-5: 1.
The invention also relates to a method for preparing the monoclonal antibody drug oral nanogel, which comprises the following steps:
(1) synthesis of the crosslinking agent: weighing a proper amount of 2-hydroxyethyl disulfide and bis (p-nitrophenyl) carbonate, placing the mixture into a round-bottom flask, adding anhydrous acetonitrile, stirring for dissolving, dropwise adding anhydrous triethylamine, stirring for reacting for 18-24 hours at room temperature, after the reaction is finished, carrying out reduced pressure rotary evaporation on the reaction liquid to completely spin the solvent, adding a proper amount of dichloromethane, washing with pure water, drying with anhydrous sodium sulfate, concentrating, precipitating with a mixed solution of ethyl acetate and petroleum ether, washing until no impurity exists, obtaining a light yellow solid, and drying to obtain the disulfide bond cross-linking agent.
(2) Preparation of nanogel: completely dissolving an antibody by using a buffer solution with the pH of 7.4 to obtain a monoclonal antibody solution, weighing a cross-linking agent, dissolving by using DMSO (dimethyl sulfoxide), slowly dropwise adding the monoclonal antibody solution, and reacting at the temperature of 20-30 ℃ for 20-30 minutes to obtain a reaction solution; and dropwise adding a hyaluronic acid solution into the reaction solution, continuously reacting for 20-30 minutes at 20-30 ℃, dialyzing to remove unreacted impurities, and removing unreacted monoclonal antibodies by using a sephadex column to obtain the monoclonal antibody medicine oral nanogel, wherein the particle size of the nanogel is 100-200 nm.
Preferably, the ratio of the amount of hyaluronic acid to the amount of monoclonal antibody substance in step (2) is 1-5: 1. The ratio of the amount of the cross-linking agent to the amount of the monoclonal antibody substance in the step (2) is 10-30: 1.
Preferably, the monoclonal antibody is infliximab, adalimumab or certolizumab ozogamicin. The monoclonal antibody can be replaced by other antibody medicines, water-soluble proteins and polypeptides with immunoregulation or anti-inflammatory properties
The invention has the following beneficial effects: (1) the monoclonal antibody medicine oral nano gel can directly transport the antibody medicine to an inflammation part by adopting an oral administration means, can avoid adverse reaction caused by systemic administration of the medicine, and simultaneously increases the compliance of patients. (2) The oral nanogel prepared by the method can improve the drug loading rate and the encapsulation rate of the antibody drug, and the adopted preparation process can fully ensure the activity of the protein drug. (3) The nanogel prepared by the method can be gathered at the inflammatory part of the intestinal tract through a targeting effect, and simultaneously, the S-S bond crosslinked by the antibody can be degraded in a responsive manner under the intestinal tract environment to release the drug, so that the targeted and positioned release of the drug at the inflammatory part of the intestinal tract is realized.
(IV) description of the drawings
FIG. 1 is a synthetic route for a crosslinker.
Fig. 2 is a TEM image of adalimumab nanogel.
Figure 3 is a graph of adalimumab gel release over time. (A) Free adalimumab bands; (B) adalimumab nanogel; (C-I) 0.5h, 1h, 2h, 4h, 8h, 12h and 24h of adalimumab release strip.
FIG. 4 shows the colon length after administration to mice of different administration groups.
(V) detailed description of the preferred embodiments
The present invention is further illustrated by the following examples, without limiting the scope of the invention thereto.
Example 1:
this example illustrates the preparation of adalimumab oral nanogels using the methods provided by the present invention.
The preparation method comprises the following steps:
1. synthesis of the crosslinking agent
Weighing appropriate amount of 2-hydroxyethyl disulfide and di (p-nitrophenyl) carbonate (the amount of substances is 1:2.4), placing the mixture in a round-bottom flask, adding 10mL of anhydrous acetonitrile, stirring to dissolve the acetonitrile, dropwise adding anhydrous triethylamine (the amount of substances is 2 times of that of the 2-hydroxyethyl disulfide), and stirring to react at room temperature for 24 hours. After the reaction is finished, carrying out reduced pressure rotary evaporation on the reaction liquid at the temperature of 45 ℃ to completely dry the solvent, adding a proper amount of dichloromethane, washing for 3 times by pure water, drying by anhydrous sodium sulfate, concentrating, precipitating by ethyl acetate and petroleum ether in a ratio of 1:3, washing until no impurities exist to obtain a light yellow solid, and drying to obtain light yellow solid powder, namely the cross-linking agent. See FIG. 1 for synthetic and other routes.
2. Preparation of nanogels
1mg of the crosslinking agent was weighed out and dissolved in 1mL of DMSO. 1mL of adalimumab-containing PBS solution at a concentration of 0.1. mu. mol/mL was taken, and the pH was adjusted to 7.4. The cross-linking agent in DMSO solution was slowly added dropwise to the antibody solution, and the reaction was carried out at 25 ℃ for 30 minutes. 1mL of a 0.2. mu. mol/mL solution of hyaluronic acid in PBS was added dropwise to the reaction solution, and the reaction was continued at 25 ℃ for 30 minutes. Dialyzing to remove unreacted impurities, and finally removing the impurities by using a Sephadex G100 column to obtain adalimumab nanogel with the drug loading rate of 56.8 percent and the encapsulation rate of 98.5 percent, wherein a TEM image of the adalimumab nanogel is shown in figure 2.
3. Nanogel in vitro release assay
To detect the responsive release behavior of the nanogels, the antibody nanogels were incubated with a phosphonium salt. The phosphonium salt can specifically cleave the disulfide bonds, converting the antibody gel to the free antibody form. The antibody gel was placed in a phosphate buffer at pH 7.4 and shaken at a constant temperature of 100r/min at 37 ℃ for the release test. At predetermined time points (0.5h, 1h, 2h, 4h, 8h, 12h, 24h), 100 μ L of supernatant was taken, run on gel, and used for drug release assay. The release assay was performed in triplicate and the results are shown in figure 3.
As can be seen, the drug of the nanogel is continuously released under the action of the phosphonium salt, less and less gel is left in the sample loading hole, and more free antibodies are left, which indicates that the nanogel can be reduced into the free antibodies in the intestinal tract.
4. Pharmacodynamic experiment
A mouse model of enteritis was made using TNBS. Enteritis mice were randomly divided into 3 groups, and the results were shown in fig. 4, in which gastric physiological saline (control group), adalimumab group (5mg/kg) was administered to the tail vein, adalimumab group (5mg/kg) was administered orally, the administration was continued for three days, and the mice were sacrificed 5 days after molding, and intestinal tissues were taken out and measured for length.
The results show that the intestinal length of mice in the oral nanogel administration group is closer to that of mice in the healthy group than that in the tail vein administration group, which indicates that the intestinal length can be better retained by the oral nanogel, and the oral nanogel shows excellent pharmacodynamic properties.
Example 2:
the example is the adalimumab oral nano preparation prepared by the method provided by the invention.
The preparation method comprises the following steps:
1. synthesis of the crosslinking agent
The same as in example 1.
2. Preparation of nanogels
1mg of the crosslinking agent was weighed out and dissolved in 1mL of DMSO. 1mL of adalimumab-containing PBS solution at a concentration of 0.08. mu. mol/mL was taken, and the pH was adjusted to 7.4. The cross-linking agent in DMSO solution was slowly added dropwise to the antibody solution, and the reaction was carried out at 25 ℃ for 30 minutes. 1mL of a 0.16. mu. mol/mL solution of hyaluronic acid in PBS was added dropwise to the reaction solution, and the reaction was continued at 25 ℃ for 30 minutes. Dialyzing to remove unreacted impurities, and finally removing the impurities by using a Sephadex G100 column to obtain the adalimumab nanogel with the drug loading rate of 56.6 percent and the encapsulation rate of 99 percent.
Example 3:
the example is the adalimumab oral nano preparation prepared by the method provided by the invention.
The preparation method comprises the following steps:
1. synthesis of the crosslinking agent
The same as in example 1.
2. Preparation of nanogels
1mg of the crosslinking agent was weighed out and dissolved in 1mL of DMSO. 1mL of adalimumab-containing PBS solution at a concentration of 0.13. mu. mol/mL was taken, and the pH was adjusted to 7.4. The cross-linking agent in DMSO solution was slowly added dropwise to the antibody solution, and the reaction was carried out at 25 ℃ for 30 minutes. 1mL of a 0.26. mu. mol/mL solution of hyaluronic acid in PBS was added dropwise to the reaction solution, and the reaction was continued at 25 ℃ for 30 minutes. Dialyzing to remove unreacted impurities, and finally removing the impurities by using a Sephadex G100 column to obtain the adalimumab nanogel with the drug-loading rate of 53.8 percent and the encapsulation rate of 92.5 percent.
While the preferred embodiments and principles of this invention have been described in detail, it will be apparent to those skilled in the art that variations may be made in the embodiments based on the teachings of the invention and such variations are considered to be within the scope of the invention.
Claims (7)
1. An oral nanogel of a monoclonal antibody medicament is prepared by the following method: and (2) crosslinking the mono-antibody medicament by using a disulfide bond crosslinking agent (2- [ (2- { [ (4-nitrophenoxy) carbonyl ] oxy } ethyl) disulfide group ] ethyl 4-nitrobenzoate) or a homolog thereof to form a nano-crosslinked substance, and then connecting hyaluronic acid to the surface of the nano-crosslinked substance through chemical synthesis to obtain the mono-antibody medicament oral nanogel.
2. The monoclonal antibody-based drug oral nanogel of claim 1, wherein the monoclonal antibody drug is an anti-TNF- α monoclonal antibody drug.
3. The monoclonal antibody drug oral nanogel according to claim 1, wherein the hyaluronic acid is an aminated hyaluronic acid with a molecular weight ranging from 10 to 100kDa, and the ratio of the hyaluronic acid to the amount of the monoclonal antibody drug substance is 1-5: 1.
4. A method of preparing a monoclonal antibody-based pharmaceutical oral nanogel according to claim 1, comprising the steps of:
(1) synthesis of the crosslinking agent: weighing a proper amount of 2-hydroxyethyl disulfide and bis (p-nitrophenyl) carbonate, placing the mixture into a round-bottom flask, adding anhydrous acetonitrile, stirring for dissolving, dropwise adding anhydrous triethylamine, stirring for reacting for 18-24 hours at room temperature, after the reaction is finished, carrying out reduced pressure rotary evaporation on the reaction liquid to completely spin the solvent, adding a proper amount of dichloromethane, washing with pure water, drying with anhydrous sodium sulfate, concentrating, precipitating with a mixed solution of ethyl acetate and petroleum ether, washing until no impurity exists to obtain a light yellow solid, and drying to obtain a disulfide bond cross-linking agent (2- [ (2- { [ (4-nitrophenoxy) carbonyl ] oxy } ethyl) disulfide ] ethyl 4-nitrophenylcarbonate);
(2) preparation of nanogel: completely dissolving an antibody by using a buffer solution with the pH of 7.4 to obtain a monoclonal antibody solution, weighing a cross-linking agent, dissolving by using DMSO (dimethyl sulfoxide), slowly dropwise adding the monoclonal antibody solution, and reacting at the temperature of 20-30 ℃ for 20-30 minutes to obtain a reaction solution; and dropwise adding a hyaluronic acid solution into the reaction solution, continuously reacting for 20-30 minutes at 20-30 ℃, dialyzing to remove unreacted impurities, and removing unreacted monoclonal antibodies by using a sephadex column to obtain the monoclonal antibody medicine oral nanogel.
5. The method of claim 4, wherein the amount of hyaluronic acid to monoclonal antibody in step (2) is 1-5: 1.
6. The method of claim 4, wherein the mass ratio of the cross-linking agent to the monoclonal antibody in step (2) is 10-30: 1.
7. The method of claim 4, wherein the monoclonal antibody is infliximab, adalimumab, or certolizumab ozogamicin.
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Cited By (2)
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| CN114404354A (en) * | 2021-11-12 | 2022-04-29 | 合肥启灏医疗科技有限公司 | Nasal in situ gel preparation and preparation method thereof |
| CN115581768A (en) * | 2022-08-31 | 2023-01-10 | 浙江大学医学院附属第一医院 | ROCKI oral nano preparation for resisting intestinal fibrosis and preparation method thereof |
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