JP3018463B2 - Glucose-responsive drug-releasing complex - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a glucose-responsive drug release complex.

The complex is a substance that releases a required amount of a drug in accordance with the glucose concentration in the environment, and is desired to be taken in response to an increase in blood glucose level after eating, such as insulin administration in diabetes. It is used for drug administration and used as various preparations.

(Prior Art) In a living body, in a healthy state, the in-vivo regulation mechanism (homeostasis) works sufficiently, and for example, various kinds of ions such as ion concentration and blood glucose level in various bloods are always kept constant. Tightly adjusted by the advanced feedback system. However, when a problem occurs in this regulatory mechanism for some reason, for example, when a chronic illness such as diabetes or hypertension occurs, insulin or a drug as an external therapeutic agent is periodically administered according to the symptoms. Need to be administered. The amount and timing of the drug to be administered at that time should be carefully considered.
In particular, a great change in the balance of the regulatory mechanism in the living body has serious consequences, and thus requires special care.

From this point of view, there has been a lot of interest in the study of formulations with an auto-feedback mechanism that releases the drug only when necessary and stops the release immediately when it is normal, that is, intelligent formulations, and many studies have been made. .

Elliott and his colleagues detect blood glucose with a glucose detector,
A small portable device that injects a corresponding amount of insulin into a vein using a pump has been reported (J.Am.Me
d. Assoc., 241 , 223 (1979)).

Also, S. W. Kim et al. Have presented an insulin release system using a complex of concanavalin A and a sugar chain-modified insulin as a molecular device having both a glucose sensor function and a drug release function (DI
ABETES, 32, 499 (1983) ).

(Problems to be Solved by the Invention) In the example of Elliott et al., A blood sugar level sensor is installed in a bloodstream, while an insulin infusion pump, a control circuit, and a power supply are housed in a cigarette box-sized case, and the blood glucose sensor is placed outside a living body. It is fixed with a belt or the like and always carried and used. However, in this case, since the glucose sensor is connected to the patient's bloodstream through the skin for a long time, there are problems such as bacterial infection and thrombus generation from the connection port, and furthermore,
Safety and reliability are not sufficient because the injection needle is likely to be clogged due to crystallization of insulin, and trouble due to mechanical or electronic circuit failure is expected.

The example of Kim et al. Is prepared by dispersing the above-mentioned complex in a pouch made of a polymer film. If this is implanted in the abdominal cavity, an exchange reaction occurs between the sugar chain-modified insulin bound to concanavalin A and glucose with an increase in the glucose concentration outside the pouch,
Glycosylated insulin is released. On the other hand, the system has an auto-feedback mechanism in which when the glucose concentration decreases, the exchange reaction decreases and the insulin release also decreases. However, this system uses concanavalin A, which is extremely toxic, and thus has a problem in safety as a drug.

As described above, it is desired that the glucose concentration-responsive drug releasing substance has both the glucose sensing function and the drug releasing function in a single molecular device. Further, in consideration of use as a drug, the substance must have extremely low toxicity and must be a substance that can be formed into various dosage forms.

An object of the present invention is to provide a glucose-responsive drug release complex that is excellent in selective response to glucose, has low toxicity, and can be used as an intelligent preparation that can be formed into various dosage forms. I have.

(Means for Solving the Problems) The present invention relates to an activity in which a benzeneboronic acid derivative immobilized on a carrier and a drug having a cis-diol group or a polyvalent hydroxyl group equivalent thereto are bonded via a boronate ester bond. The present invention provides a glucose-responsive drug-releasing complex having a site.

 The composite of the present invention is represented by the following general formula.

M: carrier Curve: arm containing covalent bond, ionic bond, Schiff base, etc. X: substituents such as H, NO ₂ , SOCH ₃ , SO ₂ CH ₃ , SOCF ₃ , SO ₂ CF _3, etc. A drug having a diol group or a polyvalent hydroxyl group equivalent thereto. More specifically, the substance is in vivo, in vitro, ix vivo, as shown in the following formula.
Depending on the glucose concentration in the environment, a substitution reaction occurs between glucose and a drug having a cis-diol group or a polyvalent hydroxyl group equivalent thereto at the boronic acid ester binding site of the substance, whereby the drug Is a glucose-responsive drug releasing substance that controls the release of

Drugs applicable to the substance include, for example,
Sugar chain-modified protein drugs such as glucosylated insulin derivatives disclosed in 502065, or bronchodilator isoproterenol, isoetalin,
Epirin, butaneefylin, WG253, trimethokinol and the like, or chitokin and the like inotropic drugs, or kanamycin, anti-tuberculosis drugs, ribidomycin, streptomycin and the like, such as cis-diol group in the skeleton or Any drug having a polyvalent hydroxyl group equivalent thereto may be used, and two or more drugs may be used in combination.

The benzeneboronic acid derivative-supporting substance used in the complex of the present invention includes a commercially available packing material for an affinity chromatography column containing 3-aminobenzeneboronic acid, for example, the product name of Matrex (registered trademark) PBA series manufactured by Amicon or And Affigel 601 manufactured by Bio-Rad Co., Ltd. can be used.

Further, it can also be prepared using a known general polymer synthesis method or polymer modification method.

For example, Japanese Patent Application Laid-Open No. 62-192405 discloses that a vinyl alcohol copolymer gel is first treated with epichlorohydrin to introduce an epoxy group, and then further treated with 3-aminobenzeneboronic acid hemisulfate to form 0.28. It is described that a polyvinyl alcohol copolymer gel having a 3-aminobenzeneboronic acid content of meq / g is obtained. Also, as another method, a diameter of 10μ
m is treated with a silane coupling agent having a glycidyl group at the end, and then reacted with 3-aminobenzeneboronic acid to obtain silica gel carrying 3-aminobenzeneboronic acid (M. Akashi, Nucleic
Acids Symp. Ser., 16 , 41 (1985)).

Further, for example, it can also be obtained by copolymerizing a vinyl derivative of benzeneboronic acid such as N- (meth) acryloylaminobenzeneboronic acid or vinylbenzeneboronic acid with another copolymerizable comonomer. Here, the monovalent comonomer is acrylamide,
N-methylacrylamide, N, N-dimethylacrylamide, besides N, N-dimethylaminopropylacrylamide, N, N-dimethylaminoethyl acrylate and various quaternary salts, further acrylic acid, various alkyl acrylates, methacrylic acid, various alkyls Methacrylate, hydroxyethyl methacrylate, N-vinylpyrrolidone, acryloylmorpholine, acrylonitrile, styrene, various macromonomers, etc., and divalent comonomers include N, N'-methylene-bis (acrylamide), ethylene glycol dimethacrylate, allyl methacrylate 1 selected from allyl acrylate, etc.
Species or two or more species. Examples of the polymerization initiator include benzoyl peroxide, diisopropyl peroxydicarbonate, tertiary butyl peroxy-2-ethylhexanoate, tertiary butyl peroxypivalate, tertiary butyl peroxydiisobutyrate, lauroyl peroxide, azobis Isobutyronitrile,
Alternatively, one or more selected from the group of various redox initiator systems can be used.

For example, by polymerizing 3-methacryloylaminobenzeneboronic acid or 4-vinylbenzeneboronic acid with acrylamide and N, N'-methylene-bis (acrylamide) in an aqueous solution in the presence of a suitable initiator, benzeneboron can be obtained. An acid group-containing hydrogel can be obtained. Further, a water-soluble polymer containing a benzeneboronic acid group is obtained by copolymerizing a water-soluble comonomer such as acrylamide or N-vinylpyrrolidone with 3-methacryloylaminobenzeneboronic acid or 4-vinylbenzeneboronic acid. Can be. Similarly, a fat-soluble polymer containing a benzeneboronic acid group can be obtained by copolymerizing a fat-soluble comonomer such as stearyl methacrylate.

Next, the various benzeneboronic acid group-containing substances thus obtained and one or more of the above-mentioned drugs having a cis-diol group or a polyvalent hydroxyl group equivalent thereto are appropriately buffered. In solution, 0-100 ° C, 1 minute-2
By reacting for 4 hours, a boronic ester bond is formed between the dihydroxyboronyl group in benzeneboronic acid and the cis-diol group or the equivalent polyvalent hydroxyl group in the skeleton of the drug, so that it can be easily formed. The composite of the present invention can be obtained. In addition, as a buffer used for this reaction, a sodium phosphate buffer, a sodium acetate buffer, an o-chlorophenol buffer, a sodium arsenate buffer, a sodium carbonate buffer, a Hepes buffer, and the like can be used. The pH value at this time is 6.0 to 10.0.
0 is sometimes preferred.

On the other hand, depending on the type of drug used, it is also possible to synthesize the complex of the drug and the benzeneboronic acid derivative in advance, and then subject it to the above-described polymer synthesis reaction or polymer modification method to prepare the complex of the present invention. it can.

As described above, the complex of the present invention can obtain various forms of substances such as an inorganic carrier, a crosslinkable polymer, a natural polymer, a hydrogel, a water-soluble polymer, and a fat-soluble polymer by various preparation methods. Can be. Therefore, the complex of the present invention can be widely applied as various preparations suitable for administration methods, and can be used in various dosage forms such as injection solutions and tablets.
Can be used for Vivo, In Vitro, and Ix Vivo.

(Effect of the Invention) In the complex of the present invention, as the glucose concentration in the environment increases, an exchange reaction occurs between the drug bound to benzeneboronic acid and glucose, and the drug is released, while the glucose concentration decreases. This system has an auto-feedback mechanism in which the exchange reaction decreases as the amount decreases, and the drug release amount also decreases.

That is, the complex of the present invention controls the release of the drug in response to the glucose concentration in the environment, has low toxicity, and can be prepared in various forms. It is a substance with excellent safety and easy handling.

For example, as a signal produced by a living body, a sharp increase in blood glucose level occurring around 30 minutes after a meal is remarkable, but in response to this, an intelligent drug that releases a drug is administered to a diabetic patient after insulin administration or after a meal. It is effective for administration of a drug that is desired to be taken. Therefore, there is no need to repeat injections and the like every time for patients who require long-term administration, and it is possible to reduce the complexity and pain for doctors and patients.

(Examples) Next, the present invention will be described in more detail with reference to Examples.
The present invention is not limited to these.

First, a general procedure for evaluating glucose-responsive drug release in vitro using a column will be described.

Approximately 2 m of a commercially available or self-synthesized column packing material supporting benzeneboronic acid was packed in a polypropylene column having a diameter of 10 mm and a length of 50 mm to form a natural dropping type flow path. The effluent is a UV detector (280n
m). The packed column is filled with Hepes buffer (pH
8.5 or 7.0), then 0.1 ~ 1g / m
After about 2 m of the drug-Hepes buffer (pH 8.5 or 7.0) was injected and developed, the mixture was incubated for 30 minutes at room temperature. Then, wash thoroughly with Hepes buffer (pH 8.5 or 7.0) and confirm the stability of the baseline.
It was converted to a Hepes buffer solution containing 0 mg / dl glucose (pH 8.5 or 7.0; manufactured by Wako Pure Chemical Industries, Ltd.), and the response of drug release was evaluated.

 The results are shown in Table 1.

Example 1 Hepes buffer (pH 8.5) was used as a buffer, and Matrex (registered trademark) PBA-30 (benzeneboronic acid-crosslinked agarose gel manufactured by Amicon) was used as a substance containing a benzeneboronic acid group. Ligand concentration of 30-50 μmol boron / m gel).
Glucosylated insulin represented by the following formula described in -502065 was used. Note that bovine insulin manufactured by Sigma was used as insulin.

In the formula, m is an integer of 1 to 3. Example 2 Hepes buffer (pH 8.5) was used as a buffer, and Matrex (registered trademark) PBA- manufactured by Amicon was used as a substance containing a benzeneboronic acid group. 30 (benzeneboronic acid-crosslinked agarose gel, ligand concentration 30-50 μmol boron / m gel), and as a drug,
Glucosylated insulin represented by the following formula described in -502065 was used. Note that bovine insulin manufactured by Sigma was used as insulin.

In the formula, n is an integer of 1 to 3. Example 3 Hepes buffer (pH 8.5) was used as a buffer, and 3-methacryloylaminobenzeneboronic acid / acrylamide / bisacrylamide was used as a substance containing a benzeneboronic acid group. 5 / 94.5 / 0.5 mol / mol / mol) copolymerized beaded hydrogel (about 30 μm in diameter)
As a drug, bronchodilator isoprotenol was used.

Example 4 Hepes buffer (pH 8.5) was used as a buffer, and 3-methacryloylaminobenzeneboronic acid / acrylamide / bisacrylamide (5 / 94.5 / 0.5 mol / mol / mol) was used as a substance containing a benzeneboronic acid group. A) hydrogel (diameter of about 30 μm) prepared by copolymerization of
The drug used was ribidomycin, an antituberculous drug.

Example 5 Hepes buffer (pH 7.0) was used as a buffer, and a substance containing a benzeneboronic acid group was described in a literature (M. Akashi, Nuc).
Leic Acids Symp. Ser., 16 , 41 (1985)). Silica gel beads (about 10 μm in diameter) for column packing supporting nitrated aminobenzeneboronic acid prepared by the method described in Leic Acids Symp.
And the glucosylated insulin used in Example 1 as a drug.

Example 6 Hepes buffer (pH 7.0) was used as a buffer, and a substance containing a benzeneboronic acid group was described in a literature (M. Akashi, Nuc).
Leic Acids Symp. Ser., 16 , 41 (1985)). Silica gel beads (about 10 μm in diameter) for column packing supporting nitrated aminobenzeneboronic acid prepared by the method described in Leic Acids Symp.
And the glucosylated insulin used in Example 2 as a drug.

Example 7 A Hepes buffer (pH 8.5) was used as a buffer, and a substance containing a benzeneboronic acid group was described in the literature (SABarker,
BMHatt, PJSomers, RRWoodbury, Carbohydrate Rase
arch, 26 , 55 (1973)).
Resin (60-120 mesh) synthesized by copolymerization of vinylbenzeneboronic acid / ethylvinylbenzene / divinylbenzene (95.7 / 2.0 / 2.3 mol / mol / mol), and glucosyl used as a drug in Example 2 Insulin was used.

Example 8 Hepes buffer (pH 8.5) was used as a buffer, and 3- (methacryloylaminobenzeneboronic acid / acrylamide / ethylenebisacrylamide (4/90/6 mol / mol / mol / mol / mol / mol) was used as a substance containing a benzeneboronic acid group. mol), a beaded hydrogel (diameter of about 300 μm, ligand (boronic acid) concentration of 20 to 30 μmol / polymer m, degree of swelling of about 9 g / g of polymer) prepared by copolymerization, and a drug represented by the following formula Polyhydric hydroxyl-modified insulin was used.
Chemical modification of insulin includes Sigma bovine insulin,
DSS (disuccinimidyl subvalate) manufactured by Pierce Co., Ltd. was used as a crosslinking agent, and tris (hydroxymethyl) aminomethane manufactured by Tokyo Chemical Industry Co., Ltd. was used as a polyvalent hydroxyl compound.

In the formula, n is an integer of 1 to 3 Examples 1 to 8 are summarized in Table 1 for the drug release amount.

Hepes buffer containing glucose as eluent, A: 10
0 mg / dl, B: 200 mg / dl, and C: 300 mg / dl, respectively. For D, a Hepes buffer containing no glucose was used as an eluate.

[Brief description of the drawings]

FIG. 1 shows a chromatograph obtained in Example 1 as a representative example of each Example. The arrows in the figure are A; Hepes buffer containing 100 mg / dl glucose as eluent, B;
Hepes buffer containing 200 mg / dl glucose, C; also 30
This indicates that a Hepes buffer containing 0 mg / dl of glucose was supplied.

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Mitsuo Okano, Inventor 6-12-12, Kokufudai, Ichikawa, Chiba Prefecture (72) Inventor, Kazunori Kataoka, 1083-4, Omuro, Kashiwa, Chiba Prefecture (72) Inventor, Yoshiyuki Koyama Noda, Chiba Prefecture 2-42-26 Iwana, Ichikawa (72) Inventor Masayuki Yokoyama 5-26-25, Higashioi, Shinagawa-ku, Tokyo (56) References JP-A-4-244031 (JP, A) , A)

Claims (1)

(57) [Claims] (1) An active site in which a benzeneboronic acid derivative immobilized on a carrier and a drug having a cis-diol group or a polyvalent hydroxyl group equivalent thereto are bonded via a boronate ester bond. Glucose responsive drug release complex.