WO2012146110A1

WO2012146110A1 - Insulin-delivering transdermal formulation and preparation method therefor

Info

Publication number: WO2012146110A1
Application number: PCT/CN2012/073017
Authority: WO
Inventors: 王义明
Original assignee: Wang Yiming
Priority date: 2011-04-27
Filing date: 2012-03-26
Publication date: 2012-11-01
Also published as: CN102178662B; CN102178662A

Abstract

A insulin-delivering transdermal formulation and preparation method therefor. The insulin-delivering transdermal formulation comprises insulin, phospholipid, cholesterol, a surface active agent, a transdermal penetration enhancer, triethanolamine in hydrochloric acid and gel. The insulin-delivering transdermal formulation prepared according to the present invention has a uniform particle size, high encapsulation rate, high stability, is convenient to administrate, and improves patient compliance.

Description

Transdermal delivery preparation for insulin delivery body and preparation method thereof

The present invention relates to a transdermal delivery preparation for an insulin delivery body and a preparation method thereof. Background technique

Insulin is the drug of choice for the treatment of diabetes. It is a protein consisting of 51 amino acids. The tertiary end of the structure is easy to form stable non-parallel and covalent β-sheets, forming a dimer with another molecule of insulin, and in zinc. In the presence of ions, the three dimers can be combined into a more stable hexamer. This polymer has a large molecular weight, is difficult to penetrate the absorption barrier, and is easily degraded by gastric acid and gastrointestinal digestive enzymes. Therefore, the mode of administration of insulin has always been injectable, and long-term injection administration causes great pain and inconvenience to patients. There may even be adverse reactions such as allergies, induration, inflammation, etc. The study is safe and effective, and improves the bioavailability of insulin in the non-injection route, which has been the focus of the medical and pharmaceutical circles at home and abroad.

In recent years, oral insulin and inhaled insulin have also been hot topics in pharmaceutics research. However, the problems faced by oral administration are: (1) decomposition of proteolytic enzymes in the digestive tract; (2) large insulin molecules, easy to polymerize; (3) difficult to absorb and the first pass effect of the liver. The problems faced by inhaled insulin are: Potentially dangerous to the lungs, especially to the fragile alveolar/capillary structures.

Therefore, transdermal administration is an ideal method of administration of insulin. The hydrolase activity of the skin is quite small, which is beneficial to the stability of the polypeptide and protein drugs. Compared with the injection, the transdermal administration can maintain a constant blood concentration, can be administered autonomously, is safe and convenient, and can avoid the liver. Over-effect and degradation of the gastrointestinal tract. But the biggest problem is that the penetration of insulin drugs into the skin is too weak. Currently, methods for increasing skin permeability are mainly applied by penetration enhancers, iontophoresis, ultrasonic waves, and electroporation methods. A carrier can also be applied to increase the transdermal efficiency of the drug. Summary of the invention

In order to solve the above technical problems, the present invention provides a transdermal administration preparation for an insulin delivery body and a preparation method thereof.

The transdermal delivery preparation of the insulin delivery body provided by the present invention contains the following components: insulin, phospholipid, cholesterol, surfactant, transdermal penetration enhancer, triethanolamine-hydrochloric acid and gel.

Preferably, the above-mentioned transdermal delivery preparation of insulin delivery body contains the following components by weight: insulin 0.2-1.0; 5-30;

1-5;

Ethanolamine-hydrochloric acid buffer 50-80;

1-5.

The transdermal delivery preparation of the above insulin delivery body further contains ethanol.

Preferably, the above phospholipid is dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, distearoylphosphatidylethanolamine or soybean phosphatidylcholine; the above surfactant is sodium cholate or deoxycholic acid Sodium; the above transdermal penetration enhancer is one of propylene glycol, ethanol, azone, eucalyptus oil, D-lemon, L-menthol, turpentine, camphor oil and geranium oil, or any combination of two or more thereof; The gel is xanthan gum, carbomer or poloxamer 188. More preferably, the phospholipid is soybean phosphatidylcholine, the surfactant is sodium deoxycholate, the transdermal penetration enhancer is eucalyptus oil, and the gel is carbomer.

Further preferably, the above-mentioned transdermally administered preparation of insulin delivery body contains the following components by weight: insulin 8.0;

Soy phosphatidylcholine 8.0;

Cholesterol 8.0;

Sodium deoxycholate 1.0;

Eucalyptus oil 1.0;

Triethanolamine-hydrochloric acid buffer 60 having a pH of 6.0 6.8;

Carbomer 1.

The above transdermal delivery preparation for insulin delivery body also includes ethanol.

The present invention also provides a method for preparing the above-mentioned insulin delivery body gel transdermal administration preparation, and the steps are as follows:

(1) taking phospholipids, cholesterol, surfactants and transdermal penetration enhancers, dissolved in ethanol;

(2) taking insulin, dissolving in triethanolamine-hydrochloric acid buffer solution, adding the obtained solution to the solution obtained in (1), homogenizing and nano-forming the liquid through a micro-jet to obtain an insulin delivery body solution;

(3) Take the gel, add water until it swells, make a gel matrix, slowly add the insulin delivery solution of step (2) to it, and stir it evenly.

The invention can achieve the following technical effects:

1. Preparation of a transdermal delivery preparation of insulin using a transfersome. The transfersome is added to the phospholipid component of common liposomes. The addition of a surfactant such as sodium cholate gives the liposome a high degree of deformability. By changing the physical properties of the drug molecule, the osmotic pressure difference is used as a driving force to force the insulin macromolecule to deform, and smoothly pass through the micropores on the skin much smaller than the transfer body, thereby promoting transdermal absorption of the drug. The mechanism for promoting permeation is: Sodium cholate can be inserted into the bilayer of phospholipids, the distance between phospholipid molecules is increased, and the order of phosphatidyl chains is disturbed, thereby enhancing fluidity. The flexible liposome has a strong skin penetration effect and can be used as an effective carrier for transdermal absorption of insulin.

2, the components are combined, the prepared transdermal delivery of the insulin delivery agent has a uniform particle size (200-300 nm), high encapsulation efficiency (up to 87.2%), good stability, convenient administration, and preparation process thereof Using advanced micro-jet technology, the conditions are easy to control and suitable for mass production.

3. The transdermal delivery of the insulin delivery agent can maintain the administration time of at least 3 days, greatly shorten the administration interval and improve the patient's compliance, overcome the inconvenience caused by the traditional insulin injection and oral administration. The disadvantage that the insulin preparation is difficult to absorb in the small intestine and has low bioavailability is a good form of preparation for treating diabetes. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing blood glucose concentration and blood glucose concentration before administration of a long-acting protamine zinc insulin injection (country) and an insulin delivery agent prepared in Example 1 after transdermal administration (A), respectively. Percentage percentage (D) - time relationship diagram;

Fig. 2 is a graph showing the relationship between the blood concentration (mIU/L) and the time after the transdermal administration of the long-acting protamine zinc insulin injection (country) and the transdermal delivery preparation (A) prepared in Example 1 Figure. The present invention will be further described in conjunction with the accompanying drawings and specific embodiments, which are to be understood by those skilled in the art.

Example 1

Take soybean phosphatidylcholine 8.0g, cholesterol 8.0g, sodium deoxycholate l.Og and lmL eucalyptus oil, add ethanol, sonicate; weigh 0.2g of insulin, use 60mL of 0.01M triethanolamine-hydrochloric acid buffer (pH=6.2) Dissolve, add the solution to the solution obtained above, pass the micro-jet, 8-10 cycles per nozzle, pressure level 10 ⁴ pa, homogenize and nano-ize, get insulin transfer body solution Take lg carbomer, add appropriate amount of water to make a gel matrix, slowly add the insulin transfer body solution to it, and stir evenly.

The physical properties of the transdermal delivery preparation of the insulin delivery body prepared in this example: According to the particle size measurement, the insulin delivery body of the present embodiment was administered transdermally, and the particle size was uniform (200-300 nm); the encapsulation efficiency was 87.2% by ultracentrifugation, and the stability was good.

In vitro transdermal study of transdermal delivery formulations of the insulin delivery bodies of this example:

The experiment used a modified Franz diffusion cell (S=2.84 cm ² , V=6.5 mL) to fix the human abdominal skin between the upper and lower chambers of the Franz diffusion cell, so that the stratum corneum of the skin faces the supply pool, and 6.5 mL is added to the receiving cell. The receiving medium was magnetically stirred (600 r · mm ¹ ) in a constant temperature water bath at 32 ± 1 ° C, and 2 mL of the transdermal delivery preparation of the insulin delivery body of Example 1 was added to the supply tank, and the receiving liquid was taken out at 24 h. The receiving solution was centrifuged (10000 r · ηώ - 15 min, and the insulin content was determined by HPLC.

The results of the study showed that: the transdermal delivery preparation prepared according to Example 1 was administered 12 hours after the administration (accumulated transdermal dosage was 937 μ _§ ), which can reach the dosage required for the type of diabetic patients; The peak value (accumulated transdermal dose is 18174 μ _§ , which is the lowest dose for critically ill patients), so no toxicity is produced; the drug can be administered for at least 3 days.

Pharmacodynamic study of transdermal delivery preparations of insulin delivery bodies of this example

Healthy male mice were injected into the tail vein with a normal aqueous solution of alloxan (80 mg/kg) for 3 days, resulting in an animal model of diabetic mice. Mice in the diabetes model were divided into two groups, 6 in a group, fasted for 2 h before the experiment, and the first group was intraperitoneally injected with 10% chloral hydrate (0.1 g/mL). The abdominal hair was removed with an electric razor, placed on a holder, and 20 L of blood was taken as a blank blood sample, and 1.5 g of insulin delivery gel was uniformly applied to the skin of the mouse; the second group was given 96 IU/kg (4.4). Mg/kg) Long-acting protamine zinc insulin injection was used as a control group, and 10 L of blood was taken at 3 h, 5 h, 12 h, 24 h, 48 h, 72 h after administration, and blood glucose concentration was determined by glucose oxidase method.

10 μL of the blood sample was added to 60 μL of heparin physiological saline, centrifuged at 5000 r/min for 5 min, and plasma was prepared. The blood glucose concentration was determined by a glucose assay kit (glucose oxidase method). The measuring principle is as follows: glucose produces gluconic acid and hydrogen peroxide under the action of glucose oxidase, and hydrogen peroxide causes o-tolidine to form a blue substance under the action of peroxidase, and the colored substance is at a wavelength of 625 nm. The glucose concentration is proportional. The amount of glucose in the sample can be calculated by measuring the absorbance of the blue substance. The hypoglycemic condition after insulin administration can be reflected by the blood glucose concentration at each time after administration and the percentage of blood glucose concentration before administration, that is, D = C _t / C _Q X 100%. In the formula, the blood glucose concentration (mg/L) in mice when C _t is t; Co is the blood glucose concentration (mg/L) in mice before administration (ie, zero time). The experimental results are shown in Fig. 1. The results of the study showed that: In the diabetic mouse model, the blood glucose levels of the mice in the two groups were flat when not administered, and the blood glucose concentration of the mice was significantly reduced within 24 hours after administration of the long-acting protamine zinc insulin injection. And then returned to the level before administration; and the mice were administered the transdermal delivery preparation of the insulin delivery body of the present example. After that, the blood glucose concentration slowly decreased. The blood glucose concentration of the mice in 24 hours was significantly lower than that of the mice after the administration of the long-acting protamine zinc insulin injection. After that, the blood glucose concentration of the mice slowly rose, and there was still a certain drop at 72 hours. Blood sugar effect.

Pharmacokinetic study of transdermal delivery preparations of insulin delivery bodies of this example

SPF Kunming mice were divided into two groups, 6 were a group, fasted for 2 h before the experiment, and the first group was intraperitoneally injected with 10% chloral hydrate (0.1 g/mL). The abdominal hair was removed with an electric razor, and 1.5 g of insulin delivery gel was uniformly applied to the skin of the mouse. The second group was given 96 IU/kg (4.4 mg/kg) of long-acting protamine zinc insulin injection as a control group, respectively. 10 μL of blood was taken at lh, 2h, 4h, 8h, 12h, 24h, 36h, 48h, 60h, 72h, and the serum was pretreated, and the drug content was determined by HPLC. The insulin delivery gel and protamine were compared. Different pharmacokinetic trends for two formulations of zinc and insulin.

The results showed that the blood concentration of insulin in mice given long-acting protamine zinc insulin injection peaked at 2h, and then decreased rapidly. After 24h, the blood concentration of insulin measured in mice was low. In comparison with the detection limit; in contrast, after the transdermal administration of the insulin delivery agent, the insulin concentration in the mouse can be maintained for a certain period of time, and the elimination is slow, and can be maintained for 72 hours or more. Example 2

Take 30.0 g of distearoylphosphatidylcholine, 30.0 g of cholesterol, 5.0 g of sodium cholate, 3 mL of geranium oil and 2 ml of menthol, add 30 ml of ethanol, dissolve it by sonication; weigh 1.0 g of insulin, use 80 mL of 0.01 M triethanolamine-hydrochloric acid buffer (pH=6.0) is dissolved, the obtained solution is added to the solution obtained above, and the microjet is homogenized and nanosized to obtain an insulin delivery body solution; 5 g of xanthan gum is added, and an appropriate amount of water is added. The gel matrix is prepared, and the insulin delivery body solution is slowly added thereto, and the mixture is uniformly stirred.

The physical properties of the transdermal delivery preparation of the insulin delivery body prepared in this example:

According to the particle size measurement, the insulin delivery body of the present embodiment was transdermally administered, and the particle size was uniform (200-300 nm); the encapsulation efficiency was 85.1% by ultracentrifugation, and the stability was good.

The experiment is the same as in the first embodiment.

The results of the study showed that: the transdermal delivery preparation prepared according to Example 2, after 12 hours of administration (accumulated transdermal dosage of 901 g), can reach the dosage required for the type of diabetic patients; The peak value (accumulated transdermal dose is 16512 μ _§ , which is the lowest dose for critically ill patients), so no toxicity is produced; the drug can be administered for at least 3 days. Pharmacodynamic study of transdermal delivery preparations of insulin delivery bodies of this example

The specific steps of the experiment are the same as those in the first embodiment.

The results of the study showed that the blood glucose concentration of the insulin delivery body of the present example was slowly decreased after administration of the insulin delivery agent of the present example, and the blood glucose concentration of the mouse of 24 hours was significantly lower than that of the mouse after administration of the long-acting protamine zinc insulin injection. After the concentration, the blood glucose concentration of the mice slowly rose, and there was still a certain hypoglycemic effect at 72 hours.

Experimental specific steps are the same as in the embodiment 1

The results of the study indicate that the insulin concentration in the mouse can be maintained for a certain period of time after administration of the insulin delivery agent of the present embodiment, and the elimination is slow and can be maintained for more than 72 hours. Example 3

Take 24.0 g of distearoylphosphatidylethanolamine, 24.0 g of cholesterol, 1.7 g of sodium deoxycholate, 1.5 mL of L-menthol and 0.5 mL of turpentine, add 20 ml of ethanol, dissolve it by sonication; weigh 0.4 g of insulin. The solution was dissolved in 64 mL of 0.01 M triethanolamine-hydrochloric acid buffer (pH=6.8), and the obtained solution was added to the solution obtained above, and the microfluidization was homogenized and nanosized to obtain an insulin delivery body solution; 2.5 g of poloxamer was taken. 188, adding an appropriate amount of water to form a gel matrix, slowly adding the insulin delivery body solution thereto, and stirring uniformly.

According to the particle size measurement, the insulin delivery body of the present embodiment was administered transdermally, and the particle size was uniform (200-300 nm); the encapsulation efficiency was 86.0% by ultracentrifugation, and the stability was good.

The experiment is the same as in the first embodiment.

The results of the study showed that: the transdermal delivery preparation prepared according to Example 3, after 12 hours of administration (accumulated transdermal dosage of 921 μ _§ ), can achieve the required dosage for the type of diabetic patients; The peak value (accumulated transdermal dose is 17410 μ _§ , which is the lowest dose for critically ill patients), so no toxicity is produced; the drug can be maintained for at least 3 days.

The results of the study showed that the blood glucose concentration of the insulin delivery body of the present example was slowly decreased after administration of the insulin delivery agent of the present example, and the blood glucose concentration of the mouse of 24 hours was significantly lower than that of the mouse after administration of the long-acting protamine zinc insulin injection. After the concentration, the blood glucose concentration of the mice slowly rose, and there was still a certain hypoglycemic effect at 72 hours. Pharmacokinetic study of transdermal delivery preparations of insulin delivery bodies of this example

Experimental specific steps are the same as in the embodiment 1

The results of the study indicate that the insulin concentration in the mouse can be maintained for a certain period of time after administration of the insulin delivery agent of the present embodiment, and the elimination is slow and can be maintained for more than 72 hours. Example 4

Take dipalmitoylphosphatidylcholine 5.0g, cholesterol 2.0g, sodium deoxycholate 0.6g, 2mL camphor oil and 3mL azone, add 5ml of ethanol, soak it by ultrasound; weigh 0.2g of insulin, use 60mL 0.01 M triethanolamine-hydrochloric acid buffer (pH=6.8) dissolved, the obtained solution was added to the solution obtained above, and the micro-jet was homogenized and nano-sized to obtain an insulin delivery body solution; lg xanthan gum 188 was added, and an appropriate amount was added. The water is made into a gel matrix, and the insulin delivery body solution is slowly added thereto, and the mixture is uniformly stirred.

According to the particle size measurement, the insulin delivery body of the present embodiment was administered transdermally, and the particle size was uniform (900-1300 nm); and the encapsulation efficiency was 73.0% by ultracentrifugation.

The experiment is the same as in the first embodiment.

The results of the study showed that: the transdermal delivery preparation prepared according to Example 3, after 12 hours of administration (accumulated transdermal dosage of 794 μ _§ ), can achieve the required dosage for the type of diabetic patients; The peak value (accumulated transdermal dose is 15820 μ _§ , which is the lowest dose for critically ill patients), so no toxicity is produced; the drug can be administered for at least 3 days.

The above-described embodiments are merely preferred embodiments for fully illustrating the present invention, and the scope of protection of the present invention is not limited thereto. Equivalent substitutions or modifications made by those skilled in the art based on the present invention are within the scope of the present invention. The scope of the invention is defined by the claims.

Claims

Claim

A transdermal delivery preparation for insulin delivery, characterized by comprising the following components: insulin, phospholipid, cholesterol, surfactant, transdermal penetration enhancer, triethanolamine-hydrochloric acid and gel.

The transdermal delivery preparation for insulin delivery according to claim 1, which comprises the following components by weight:

Insulin

Lecithin

Cholesterol

Surfactant

Transdermal penetration enhancer

Ethanolamine-HCl buffer with a pH of 6.0-6.8

Gel

The transdermal delivery preparation for insulin delivery according to claim 2, which further comprises ethanol.

The transdermal delivery preparation for insulin delivery according to claim 1, wherein the phospholipid is dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, distearoylphosphatidylethanolamine Or soybean phosphatidylcholine; the surfactant is sodium cholate or sodium deoxycholate; the transdermal penetration enhancer is propylene glycol, ethanol, azone, eucalyptus oil, D-lemon, L-menthol Any one or a combination of two or more of alcohol, turpentine, camphor oil and geranium oil; the gel is xanthan gum, carbomer or poloxamer 188.

The transdermal delivery preparation for insulin delivery according to claim 4, wherein the phospholipid is soybean phosphatidylcholine, the surfactant is sodium deoxycholate, and the transdermal penetration enhances The agent is eucalyptus oil and the gel is carbomer.

The transdermal delivery preparation for insulin delivery according to claim 5, which comprises the following components by weight:

Insulin 0.2;

Soy phosphatidylcholine 8.0;

Cholesterol 8.0;

Sodium deoxycholate 1.0;

Eucalyptus oil 1.0;

Triethanolamine-hydrochloric acid buffer 60 having a pH of 6.0 6.8; Carbomer 1.

The transdermal delivery preparation for insulin delivery according to claim 6, further comprising ethanol.

The method for producing a transdermal delivery preparation for an insulin delivery body according to any one of claims 1 to 7, wherein the steps are as follows: