阅读说明：本技术 含乙烯基功能性基团的rgd序列小分子多肽、温度敏感自律式释药智能小儿退烧贴及应用 (RGD sequence micromolecule polypeptide containing vinyl functional group, temperature-sensitive self-discipline drug-release intelligent infantile fever-reducing plaster and application ) 是由 刘红 关荧莹 穆岩 秦吟 于 2020-04-28 设计创作，主要内容包括：本发明属于药物制剂技术领域,尤其涉及含乙烯基功能性基团的RGD序列小分子多肽、温度敏感自律式释药智能小儿退烧贴及应用。退烧贴由背衬层、温度敏感自律式释放退烧药凝胶系统、保护膜组成。温度敏感自律式释放退烧药凝胶系统由聚N-异丙基丙烯酰胺类水凝胶(温度敏感性凝胶)、水溶性氮酮(促渗透剂)、丙二醇(兼有增溶和保湿作用)、药物(退烧药)、蒸馏水(溶剂)组成。本发明提供的温度敏感自律式释药智能小儿退烧贴能够实现发热患儿体温在38.5℃以下时基本不释药或极其缓慢释药,而在38.5℃以上开始加速释药,可以减轻医护人员和家长频繁监测患儿体温的负担,同时还可以确保发热患儿夜间体温骤升时自动得到退烧药的及时医治。(The invention belongs to the technical field of medicinal preparations, and particularly relates to RGD sequence micromolecule polypeptide containing vinyl functional groups, a temperature-sensitive autonomous drug release intelligent infantile fever reducing plaster and application. The fever reducing plaster consists of a back lining layer, a temperature sensitive self-discipline fever reducing medicine releasing gel system and a protective film. The temperature-sensitive self-discipline antipyretic drug release gel system consists of poly N-isopropyl acrylamide hydrogel (temperature-sensitive gel), water-soluble azone (penetration enhancer), propylene glycol (having solubilization and moisturizing functions), a drug (antipyretic drug) and distilled water (solvent). The temperature-sensitive self-regulated medicine-releasing intelligent infantile fever reducing patch provided by the invention can realize that the fever of a fever infant patient is basically not released or is released extremely slowly when the body temperature is below 38.5 ℃, and the fever of the fever infant patient starts to be released at a temperature above 38.5 ℃, so that the burden of frequently monitoring the body temperature of the infant patient by medical personnel and parents can be relieved, and the fever infant patient can be automatically treated in time when the body temperature suddenly rises at night.)

1. A preparation method of RGD sequence micromolecule polypeptide containing vinyl functional groups is characterized by comprising the following steps:

1) washing and soaking the 2-chlorotrityl chloride resin in dimethylformamide;

2) enabling the aspartic acid with amino and carboxyl on the side group protected by 9-fluorenylmethyloxycarbonyl and tert-butyl to interact with the 2-chlorotrityl chloride resin in the step 1, and fixing the resin after the reaction is completed;

3) adding 20% piperidine-dimethylformamide solution into the resin obtained in the step 2, and removing a fluorenylmethyloxycarbonyl protecting group on aspartic acid;

4) adding glycine with amino protected by 9-fluorenylmethoxycarbonyl, condensing agent 1-hydroxybenzotriazole, benzotriazole-N, N, N ', N' -tetramethylurea hexafluorophosphate and catalyst N, N-diisopropylethylamine into the resin obtained in the step 3, so that C-terminal carboxyl of the glycine and N-terminal amino of the aspartic acid are fully condensed;

5) adding 20% piperidine-dimethylformamide solution into the resin obtained in the step 4, and removing a fluorenylmethyloxycarbonyl protecting group on glycine;

6) adding arginine, a condensing agent 1-hydroxybenzotriazole, benzotriazole-N, N, N ', N' -tetramethylurea hexafluorophosphate and a catalyst N, N-diisopropylethylamine, wherein the amino group and the guanidyl group on the side group are respectively protected by 9-fluorenylmethyloxycarbonyl and 2,2,4,6, 7-pentamethyl-2H-benzofuran-5-sulfonyl, into the resin obtained in the step 5, so that C-terminal carboxyl of the arginine and N-terminal amino of the glycine are fully condensed;

7) adding 20% piperidine-dimethylformamide solution into the resin obtained in the step 6, and removing a fluorenylmethyloxycarbonyl protecting group on arginine;

8) adding acrylic acid, a condensing agent 1-hydroxybenzotriazole, benzotriazole-N, N, N ', N' -tetramethylurea hexafluorophosphate and a catalyst N, N-diisopropylethylamine into the resin obtained in the step 7, and fully condensing carboxyl of the acrylic acid and N-terminal amino of arginine;

9) washing the resin obtained in step 8 with methanol and dichloromethane, and drying;

10) and (3) adding a deprotection agent trifluoroacetic acid, triisopropylsilane and distilled water into the resin obtained in the step (9), and removing tert-butyl to obtain the RGD sequence micromolecule polypeptide containing vinyl functional groups.

2. The application of the RGD sequence micromolecule polypeptide containing vinyl functional groups prepared by the preparation method of claim 1 in preparing phase-change materials.

3. A temperature-sensitive self-discipline antipyretic gel system is characterized by comprising the following components in percentage by mass:

the phase transition temperature of the gel system was 38.5 ℃.

4. The temperature-sensitive self-contained release antipyretic gel system as set forth in claim 3, wherein: the antipyretic is any one of acetaminophen, ibuprofen and indometacin.

5. The temperature-sensitive self-contained release antipyretic gel system as set forth in claim 4, wherein: the preparation method of the gel system comprises the steps of mixing the prescribed amount of propylene glycol, water-soluble azone and distilled water, sequentially adding N-isopropylacrylamide, RGD sequence micromolecule polypeptide containing vinyl functional groups, N' -methylenebisacrylamide and an ethanol-dissolved antipyretic, oscillating and mixing uniformly, introducing nitrogen, adding ammonium persulfate, and reacting at the temperature of 16-18 ℃ for 48 hours to obtain the colorless and transparent temperature-sensitive drug-loaded gel.

6. Use of the temperature sensitive self-regulated release antipyretic gel system according to any one of claims 3-5 in the preparation of a fever patch or fever reducing device.

7. A temperature-sensitive self-discipline medicine-releasing intelligent infant fever-reducing plaster is characterized in that: comprises a back lining layer, a temperature-sensitive self-regulated antipyretic gel system and a protective film.

8. The temperature-sensitive self-regulated drug release intelligent infantile fever reducing patch as claimed in claim 7, is characterized in that: the backing layer is a non-woven fabric.

9. The temperature-sensitive self-regulated drug release intelligent infantile fever reducing patch as claimed in claim 7, is characterized in that: the protective film is a PE plastic film.

10. The temperature-sensitive autonomous drug release intelligent infantile fever reducing patch according to any one of claims 7 to 9, wherein the preparation method comprises: coating the temperature sensitive medicine-carrying gel on a back lining layer, pressing a protective film, cold-pressing for molding and shearing; sterilizing with ethylene oxide, and packaging.

Technical Field

The invention belongs to the technical field of medicinal preparations, and particularly relates to RGD sequence micromolecule polypeptide containing vinyl functional groups, a temperature-sensitive autonomous drug release intelligent infantile fever reducing plaster and application.

Background

Fever, also known as fever, is a symptom that the body temperature exceeds the normal range under the action of pyrogen or central dysfunction of thermoregulation, is the most common clinical symptom and doctor-seeking reason of children, and accounts for the first place of emergency observation of children. The normal body temperature of human body is generally 36-37.3 ℃, and clinical fever is diagnosed when the body temperature exceeds 37.3 ℃, and the current experts commonly recognize the following principles about the use of drugs (antipyretics):

when the body temperature of a child is below 38.5 ℃, antipyretics are generally not recommended for the following reasons: the body is relatively less damaged at the body temperature; secondly, heating under certain conditions is possibly beneficial to enhancing the function of immune cells and improving the defense ability of the human body to infection; thirdly, the fever reducing medicine is applied too early, so that the condition of a patient is easily covered, and the diagnosis and treatment of the original disease are delayed; and fourthly, the principle of taking medicines with least or no use is followed to reduce the potential side effect possibly caused by the medicines.

When the body temperature of a child continues to exceed 38.5 ℃, antipyretics are generally recommended for the following reasons: firstly, children often suffer from damage to the central nervous system caused by headache, dysphoria, delirium or apathy, lethargy, convulsion, and the like, and are coma in severe cases and brain damage caused by overlong time; secondly, the circulatory system of the children is changed, such as symptoms of accelerated heart rate, increased oxygen consumption of cardiac muscle, increased cardiac burden, massive sweating and dehydration, and the like; ③ the metabolism of substances in the body is changed, such as the catabolism of sugar, fat and protein is obviously accelerated, the consumption of various vitamins is increased, and the substances are excessively consumed, thus leading to the emaciation and the weight reduction; fourthly, when the fever lasts, the appetite of the children patients is reduced, the digestion function is weakened, and the immunity is reduced.

Currently, antipyretics on the market are mainly administered orally and by injection. The infantile oral antipyretic mainly comprises acetaminophen (paracetamol), ibuprofen, indometacin, nimesulide and the like, and the preparation forms of the antipyretic are tablets, granules, suspensions and the like, and are often accompanied by adverse reactions such as nausea, vomiting, abdominal discomfort and the like. For children with high fever with convulsion or dysphagia, the injection of analgin, letiriin and other medicines is usually adopted clinically, which can rapidly reduce the body temperature, but will bring certain pain to the children, the compliance is poor, and simultaneously, the atrophy and spasm of the hip muscles are easily caused, and certain influence is exerted on the children nerves.

In addition, the body temperature of the fever infant needs to be continuously monitored by medical care personnel or family members due to continuous change, especially, the body temperature of the fever infant at night may rise to a high fever range or even an ultrahigh fever range due to some reason, and if the body temperature is not measured and taken in time, the physical and mental health of the infant can be greatly damaged.

Disclosure of Invention

Aiming at the problems in the prior art, the invention provides RGD sequence micromolecule polypeptide containing vinyl functional groups, a temperature-sensitive autonomous drug release intelligent infantile fever reducing plaster and application, and aims to solve part of problems in the prior art or at least alleviate part of problems in the prior art.

Aiming at the characteristics of the fever symptom of children, expert consensus and the defects of various dosage forms of the existing antipyretics, the invention provides the temperature-sensitive automatic-release intelligent infantile antipyretic patch, which can realize that the fever of a fever infant is not released basically or is released extremely slowly when the temperature is below 38.5 ℃, and the fever begins to be released at an accelerated speed when the temperature is above 38.5 ℃, can relieve the burden of medical care personnel and parents on frequently monitoring the body temperature of the infant, can also ensure that the fever infant can be automatically treated in time when the body temperature rises suddenly at night, avoids the discomfort of gastrointestinal tracts of common oral antipyretics, relieves the pain and the injury of the injection administration to the infant, and has better clinical application value and significance.

The invention is realized in such a way that a preparation method of RGD sequence micromolecule polypeptide containing vinyl functional groups comprises the following steps:

1) washing and soaking the 2-chlorotrityl chloride resin in dimethylformamide;

2) enabling the aspartic acid with amino and carboxyl on the side group protected by 9-fluorenylmethyloxycarbonyl and tert-butyl to interact with the 2-chlorotrityl chloride resin in the step 1, and fixing the resin after the reaction is completed;

3) adding 20% piperidine-dimethylformamide solution into the resin obtained in the step 2, and removing a fluorenylmethyloxycarbonyl protecting group on aspartic acid;

4) adding glycine with amino protected by 9-fluorenylmethoxycarbonyl, condensing agent 1-hydroxybenzotriazole, benzotriazole-N, N, N ', N' -tetramethylurea hexafluorophosphate and catalyst N, N-diisopropylethylamine into the resin obtained in the step 3, so that C-terminal carboxyl of the glycine and N-terminal amino of the aspartic acid are fully condensed;

5) adding 20% piperidine-dimethylformamide solution into the resin obtained in the step 4, and removing a fluorenylmethyloxycarbonyl protecting group on glycine;

6) adding arginine, a condensing agent 1-hydroxybenzotriazole, benzotriazole-N, N, N ', N' -tetramethylurea hexafluorophosphate and a catalyst N, N-diisopropylethylamine, wherein the amino group and the guanidyl group on the side group are respectively protected by 9-fluorenylmethyloxycarbonyl and 2,2,4,6, 7-pentamethyl-2H-benzofuran-5-sulfonyl, into the resin obtained in the step 5, so that C-terminal carboxyl of the arginine and N-terminal amino of the glycine are fully condensed;

7) adding 20% piperidine-dimethylformamide solution into the resin obtained in the step 6, and removing a fluorenylmethyloxycarbonyl protecting group on arginine;

8) adding acrylic acid, a condensing agent 1-hydroxybenzotriazole, benzotriazole-N, N, N ', N' -tetramethylurea hexafluorophosphate and a catalyst N, N-diisopropylethylamine into the resin obtained in the step 7, and fully condensing carboxyl of the acrylic acid and N-terminal amino of arginine;

9) washing the resin obtained in step 8 with methanol and dichloromethane, and drying;

10) and (3) adding a deprotection agent trifluoroacetic acid, triisopropylsilane and distilled water into the resin obtained in the step (9), and removing tert-butyl to obtain the RGD sequence micromolecule polypeptide containing vinyl functional groups.

The application of the RGD sequence micromolecule polypeptide containing the vinyl functional group prepared by the preparation method in preparing the phase change material.

A temperature-sensitive self-discipline antipyretic gel system comprises the following substances in percentage by mass:

the phase transition temperature of the gel system was 38.5 ℃.

The mass fractions of N-isopropylacrylamide (monomer), RGD sequence micromolecule polypeptide (monomer) containing vinyl functional group, N' -methylene-bisacrylamide (cross-linking agent) and ammonium persulfate (initiator) need to be determined by modern pharmacological means aiming at each formula (mainly determined by the type, property and content of antipyretic), so that the phase transition temperature (LCST) of the temperature-sensitive hydrogel is 38.5 ℃.

Further, the antipyretic is any one of acetaminophen, ibuprofen and indomethacin.

Further, the preparation method of the gel system comprises the steps of mixing the prescribed amount of propylene glycol, water-soluble azone and distilled water, sequentially adding N-isopropylacrylamide, RGD sequence micromolecule polypeptide containing vinyl functional groups, N' -methylenebisacrylamide and an ethanol-dissolved antipyretic, uniformly mixing by oscillation, introducing nitrogen, adding ammonium persulfate, and reacting at the temperature of 16-18 ℃ for 48 hours to obtain the colorless and transparent temperature-sensitive drug-loaded gel.

The application of the temperature-sensitive self-discipline release antipyretic gel system in preparing an antipyretic patch or an antipyretic device.

A temperature-sensitive self-regulated drug release intelligent infantile antipyretic patch comprises a back lining layer, a temperature-sensitive self-regulated drug release gel system and a protective film.

Further, the backing layer is a non-woven fabric.

Further, the protective film is a PE plastic film.

Further, the preparation method comprises the following steps: coating the temperature sensitive medicine-carrying gel on a back lining layer, pressing a protective film, cold-pressing for molding and shearing; sterilizing with ethylene oxide, and packaging.

In summary, the advantages and positive effects of the invention are:

the core technology of the invention is to synthesize hydrogel which is based on poly N-isopropylacrylamide with excellent temperature sensitivity and has the phase transition temperature (LCST) of 38.5 ℃, and prepare the intelligent infantile fever reducing plaster which can automatically turn on and off to release the fever reducing medicine at about 38.5 ℃ according to the change of body temperature by using the hydrogel.

The traditional poly-N-isopropyl acrylamide gel belongs to a thermal shrinkage type (high-temperature shrinkage type) temperature-sensitive hydrogel, the phase transition temperature (LCST) is 33 ℃, the gel is in a swelling state when the temperature is lower than the LCST, and the gel is rapidly shrunk when the temperature is higher than the LCST. In order to obtain the heat-shrinkable temperature-sensitive hydrogel with the LCST of 38.5 ℃, an inventor laboratory synthesizes an RGD sequence micromolecule polypeptide monomer containing a vinyl functional group, the RGD sequence micromolecule polypeptide in the monomer consists of arginine (Arg), glycine (Gly) and aspartic acid (Asp), and the temperature-sensitive hydrogel with higher LCST can be obtained by crosslinking and copolymerizing with an N-isopropyl acrylamide monomer through free radicals in the presence of a crosslinking agent because hydrophilic peptide bonds are used as a framework and contain a plurality of carboxyl, amino and guanidyl with good hydrophilicity and good biocompatibility. Based on the principle, the inventor optimizes the proportion of two monomers and the addition of a cross-linking agent and an initiator by using modern pharmaceutical means such as a uniform design method and the like, accurately adjusts the LCST of the temperature-sensitive gel to be higher than the expected 38.5 ℃, and well solves the key problem of the over-low LCST of the traditional poly-N-isopropylacrylamide gel system. The intelligent infant fever reducing plaster is prepared by adopting the temperature sensitive medicine-carrying hydrogel with the phase transition temperature (LCST) of 38.5 ℃, is applied to the forehead of a fever infant, when the body temperature of the infant is lower than 38.5 ℃, the temperature sensitive gel is in a swelling state, the medicine is adsorbed in a three-dimensional network of the gel and cannot be diffused out, the hydrogel plays a physical fever reducing role, when the body temperature exceeds 38.5 ℃, the gel rapidly shrinks to generate an outward discharge effect on the fever reducing medicine, the quickly released medicine enters an in-vivo blood circulation system through transdermal absorption, and the quick fever reducing effect is realized through a body temperature regulating mechanism, so that the fever reducing medicine using principle commonly recognized by current experts is met, the burden of frequently monitoring the body temperature of the infant by medical staff and parents can be relieved, and the fever reducing medicine can be automatically obtained when the body temperature of the fever infant rises at night.

Drawings

FIG. 1 is a schematic diagram of the synthesis of RGD sequence small molecule polypeptide containing vinyl functional group;

FIG. 2 is a schematic of a temperature sensitive drug loaded gel formation process;

FIG. 3 is a Differential Scanning Calorimetry (DSC) chart of the temperature sensitive drug loaded gel of example 2;

figure 4 is the in vitro cumulative drug release profile at temperature program of example 2;

FIG. 5 is a Differential Scanning Calorimetry (DSC) plot of the temperature sensitive drug loaded gel of example 3;

figure 6 is the in vitro cumulative drug release profile at temperature program of example 3;

FIG. 7 is a Differential Scanning Calorimetry (DSC) chart of the temperature sensitive drug loaded gel of example 4;

figure 8 is the in vitro cumulative drug release profile at the programmed temperature of example 4.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. The specific embodiments described herein are merely illustrative of the invention and are not intended to be limiting.

The invention discloses RGD sequence micromolecule polypeptide containing vinyl functional groups, a temperature-sensitive autonomous drug-release intelligent infantile fever reducing plaster and application thereof, which are shown in the following embodiments.