RU2385157C1 - Osteoarthritis treatment - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine and pharmacology. There is offered therapeutic agent for osteoarthritis treatment, containing hyaluronane containing 2 to 20 sugar links as an operating component. Preferentially, hyaluronane has the form of tetrasaccharide consisting of two links with a single link in the form of: -D-glucuronic acid-b-1,3-D-N-acetylglycosamine-b-1,4-(HA4). There is offered method of osteoarthritis treatment that involves the stage of introduction of effective amount of hyaluronane containing 2 to 20 sugar links to a subject requiring such treatment.

EFFECT: invention presents a new drug, effective for osteoarthritis treatment with the minimum undesired reaction.

6 cl, 1 ex, 2 tbl

Description

The technical field to which the invention relates.

The present invention relates to a therapeutic agent for the treatment of osteoarthritis, including hyaluronan as an active ingredient.

The prior art.

Hyaluronan is a long-chain polysaccharide including repeating disaccharide units of D-glucuronic acid and N-acetyl-D-glucosamine, it is also known as oligosaccharide. Hyaluronan is obtained from an extract of body tissues, such as chicken comb, umbilical cord, skin or synovial fluid, by fermentation using bacteria of the genus Streptococcus or the like. Due to the fact that it does not have a toxicological or immunological effect, hyaluronan is used in medicines or cosmetics. For example, a method for treating arthritis using intra-articular injections of hyaluronan is well known. In the description below, the hyaluronan tetrasaccharide is designated as HA4.

There is evidence that HA4 has the ability to preserve organs and treat / suppress liver diseases and stomach ulcers (patent document 1). In addition, it is also known that HA4 has the property of enhancing the expression of stress proteins and inhibiting cell death (Non-Patent Document 1).

Osteoarthritis is a joint lesion associated with chronic arthritis, in which degeneration of the components of the joint leads to destruction of the cartilage and proliferative changes in the bone and cartilage. In addition, arthritis (synovitis) develops a second time due to the above changes. In the early stages, pain occurs after overloading the joint and passes after rest. As the disease develops, pain appears even with a small load or at rest, night pains are often noted. With severe flexion or extension of the joint or with physical exertion, the joint makes a clicking sound, and synovial fluid may also accumulate in the joint that is swollen due to arthritis.

In addition to the degeneration of the components of the joint (age-related changes), the progression of the disease is associated with genetic factors, aging, obesity, joint instability, repeated subluxation and dislocation, excessive load on the joints as a result of work, sports, etc. etc.

Early treatment includes the use of anti-inflammatory or analgesic drugs and hot lotions or dressings containing an analgesic to relieve pain. In the treatment of knee joints, physical exercises aimed at strengthening the quadriceps femoris muscle are effective. Lead exercises (raising and lowering the legs) are effective in the treatment of hip joints. With osteoarthritis of the knee and hip joints, the use of a cane is also effective, since it reduces the load on the joints. Intra-articular injections of drugs protecting the joint are also effective. Caution should be taken in the use of intra-articular injectable steroid therapy, as abuse can accelerate the destruction of joints. For severe pain or degeneration, surgical operations such as osteotomy and replacement of joints with artificial joints are indicated for the treatment of knee and hip joints.

[Patent Document 1] WP 2002/004471.

[Non-patent document 1] Hu N, Ito T, Tawada A, Maeda H, Yamanokuchi H, Isahara K, Yoshida K, Uchiyama Y, Asari A. Effect of hyaluronan oligosaccharides on the expression of heat shock protein 72. J Biol Chem. 2002 10; 277 (19): 17308-14.

A brief description of the invention.

An object of the present invention is to provide a therapeutic agent for the treatment of osteoarthritis.

A subject of the present invention, a therapeutic agent for the treatment of osteoarthritis, which solves the aforementioned problem, includes hyaluronan as an active ingredient.

Since the drug of the present invention includes hyaluronan as an active ingredient, its advantage is that it can be mass-produced at a relatively low cost. In addition, taking into account that hyaluronan has practically no toxicity or antigenicity, enhances the therapeutic effect and has a preventive effect on diseases that are initially present in the body, it is assumed that it will be a therapeutic agent with a minimal undesirable drug reaction. Thus, the present invention can provide a new drug effective for the treatment of osteoarthritis.

DETAILED DESCRIPTION OF THE INVENTION

A detailed explanation of the present invention is given below. In the following explanation, the therapeutic agent of the present invention will be referred to simply as a drug.

It is sufficient that the hyaluronan contained in the drug of the present invention be in the form of a disaccharide or polysaccharide with a large number of monomer units, which mainly comprises at least one disaccharide unit, which is β-D-glucuronic acid in 1st position and β-DN-acetylglucosamine in the 3rd position, which are linked to each other, and consisted mainly of β-D-glucuronic acid and β-DN-acetylglucosamine. Hyaluronan may also be a saccharide in which these elements are associated with one or more disaccharide units, or a derivative thereof. For example, hyaluronan having a hydrolyzable protecting group such as an acyl group and the like can also be used. This saccharide may be an unsaturated saccharide, examples of unsaturated saccharides include non-reducing end saccharides, glucuronic acid, usually with an unsaturated bond between the carbon atoms in the 4th and 5th position and the like. Like the hyaluronan used in the present invention, more specifically, it is possible to use hyaluronan extracted from natural products, such as, for example, from animal products obtained by culturing microorganisms, synthesized chemically or enzymatically, and the like. So, it can be obtained from body tissues, such as chicken comb, umbilical cord, skin and synovial fluid, using known extraction and purification methods. In addition, it can also be obtained by fermentation using bacteria of the genus Streptococcus.

In the present invention, hyaluronan oligosaccharides are also within the scope of the concept of hyaluronan, and hyaluronans starting from those with a low molecular weight, such as the aforementioned disaccharide comprising one disaccharide unit and its derivatives, to high molecular weight hyaluronans having an average molecular weight can be used. weight 4,000,000. Preferred examples thereof include hyaluronans with an average molecular weight of about 380 to 900,000, which have excellent tissue penetration and the like, and more hyaluronans, including about 2 to 20 saccharide units, are descriptive.

Of the hyaluronans, it is preferable to produce those with a low molecular weight, in particular, methods aimed at reducing the molecular weight of hyaluronan using known technologies such as enzymatic cleavage, alkaline degradation, heat treatment and ultrasonic treatment (Biochem., 33 (1994) p. 6503-6507), a method for chemical or enzymatic synthesis (Glycoconjugate J., (1993) pp. 435-439; WO 93/20827) and the like. Examples of enzymatic cleavage technology include methods for producing hyaluronan oligosaccharides by exposure to hyaluronan-cleaving enzymes such as hyaluronan-cleaving enzymes (hyaluronidase derived from testes, hyaluronidase derived from Streptomyces, Acharononase anaquinone, quinonequinone, and others) chondroitinase ABC (see New Biochemistry Experiment Lecture "Carbohydrate II: Proteoglycan and glycosaminogycan," pp. 244-248, 1991, Tokyo Kagaku-dojin Publishing Company Inc.) and the like.

In addition, examples of alkaline degradation technology include a method for producing low molecular weight hyaluronan, comprising the steps of adding a base, such as about 1N sodium hydroxide, to a hyaluronan solution, heating the mixture for several hours to reduce molecular weight, and adding an acid, such as hydrochloric acid to neutralize mixtures and the like. The hyaluronan used in the present invention encompasses the salt form, and pharmacologically acceptable salts, for example, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts, such as calcium and magnesium salts, can be used to prepare the dosage form, if necessary. amine salts such as tri (n-butyl) amine salts, triethylamine salts, pyridine salts and amino acid salts and the like.

The drug of the present invention is not particularly limited, and hyaluronan with a specific molecular weight can be used alone, or hyaluronans having different molecular weights can be used in combination. The drug of the present invention includes hyaluronan as an active ingredient, and osteoarthritis can be treated by administering an effective amount of hyaluronan to mammals, including humans, without adversely affecting the body.

The medicament of the present invention can be prepared using hyaluronan or a salt thereof as such or with carriers, excipients and other additives as necessary as a medicament for oral or parenteral administration (interstitial administration, for example, intraarticular, intravenous, intramuscular or subcutaneous administration (injection), enteral administration, transdermal administration and the like), in the form of a dosage form selected without limitation, and preferably Patient-friendly, selectable without restrictions. In particular, it is preferable to prepare the drug constituting the subject of the present invention in the form of an oral dosage form.

Examples of the oral dosage form include solid dosage forms such as powders, granules, capsules and tablets, and liquid dosage forms such as syrups, elixirs and emulsions. The powder can be obtained by mixing, for example, with a filler such as lactose, starch, crystalline cellulose, calcium lactate, calcium hydrogen phosphate, magnesium aluminum metasilicate or anhydrous silicic acid. Granules can be prepared by subsequent addition, if necessary, in addition to the above excipients, for example, a binding component such as sucrose, hydroxypropyl cellulose or polyvinyl pyrrolidone, a binding component such as carboxymethyl cellulose or calcium carboxymethyl cellulose, and a disintegrant such as carboxymethyl cellulose and calcium, mixtures wet or dry. Tablets can be prepared by tableting the aforementioned powder or granules unchanged, or by adding a lubricant such as magnesium stearate or talc, and tableting the mixture. In addition, the aforementioned tablets or granules may be enteric-coated, for example with copolymers of hydroxypropyl methylcellulose phthalate and methyl methacrylate, or ethyl cellulose, or carnauba wax, hydrogenated oil or the like, to produce a intestinal disintegrating or sustained release dosage form. Hard capsules can be prepared by pouring the aforementioned powder or granules into hard capsules. Soft capsules can be prepared by mixing hyaluronan or its salt with glycerin, polyethylene glycol, sesame oil, olive oil or the like, and coating the mixture with a gelatin film. A syrup can be prepared by dissolving a sweetener in water, for example sucrose, sorbitol or glycerol and hyaluronan or a salt thereof. In addition, in addition to the sweetener and water, you can add essential oil, ethanol or the like to obtain an elixir, or Arabian gum, tragacanth gum, Polysorbate 80, sodium carboxymethyl cellulose or the like to obtain an emulsion or suspension. In addition, flavoring agents, coloring agents, preservatives, and the like can be added to these liquid dosage forms, if necessary.

Examples of the parenteral dosage form include injections, intrarectal injections, pessaries, topical transdermal agents, inhalants, aerosols, eye drops and the like. Injectable dosage forms can be prepared by adding a pH modifier to hyaluronan or a salt thereof, such as hydrochloric acid, sodium hydroxide, lactic acid, sodium lactate, monobasic sodium phosphate or sodium dihydrogen phosphate; an isotonizing agent such as sodium chloride or glucose; and distilled water for injection; sterilizing the mixture by filtering and filling the mixture into ampoules. In addition, you can get the dosage form for injection, dissolved before use, by the subsequent addition of mannitol, dextrin, cyclodextrin, gelatin or the like and freeze-drying the mixture. In addition, an emulsion for injection can also be prepared by adding an emulsifier, such as lecithin, polysorbate 80 or polyoxyethylene hydrogenated castor oil, to the hyaluronan or its salt, and emulsifying the mixture in water.

Intrarectal injections can be obtained by adding a suppository carrier, such as cocoa fatty acid mono-, di- or triglyceride or cocoa polyethylene glycol, to hyaluronan or its salt, then dissolving by heating, pouring the mixture into a mold and cooling it, or by mixing hyaluronan or its salt with polyethylene glycol, soybean oil or the like, and then coating the mixture with a gelatin film. Topical transdermal preparations can be obtained by adding white petrolatum, beeswax, liquid paraffin, polyethylene glycol or the like to hyaluronan or its salt, heating the mixture if necessary and mixing the mixture into the total mass. A dressing can be obtained by mixing the mixture into the total mass of hyaluronan or its salt with a tackifier, such as a resin or polymer of an alkyl ester of acrylic acid, and spreading the mixture onto a non-woven fabric or the like. An inhalation agent can be prepared by dissolving or dispersing hyaluronan or a salt thereof, for example, in a nebulizing agent, such as a pharmacologically acceptable inert gas, and filling the mixture in a pressure-resistant container.

Route of administration.

Methods for administering a drug of the present invention that includes hyaluronan as an active ingredient are not particularly limited, examples of which include intravenous administration, oral administration and intraarticular administration.

Dosage.

The dosage is determined appropriately depending on the disease to be treated, the patient’s age, health status and body weight and the like, and the usual daily dose is 0.05-50 mg / kg, it is taken at a time or divided into several doses.

Toxicity.

The hyaluronan used in the present invention showed little or no cytotoxicity at dosages at which biological activity as a drug was observed.

Information confirming the possibility of carrying out the invention.

The drug of the present invention will be illustrated in detail using the following examples. However, the scope of the claims of the present invention is not limited to these examples.

Example 1

In this example, to examine the effect of HA4, the drug was administered to C57BL / 6 mice, which spontaneously developed osteoarthritis.

An experimental model of osteoarthritis in an animal.

With Charles River Laboratories Japan Inc. 20-week-old male C57BL / 6 mice were purchased; animals with osteoarthritis were used. The animals were divided into a negative control group (treated with physiological saline) and a group that received HA4 orally (100 mg / 10 ml / kg).

The introduction of the test substance.

In this example, HA4 10 mg / ml was prepared. More specifically, HA4 was prepared by the method of Tawada and colleagues (Tawada A, Masa T. Oonuki Y, Watanabe A, Matsuzaki Y, Asari A. Large-scale preparation, purification, and characterization of hyaluronan oligosaccharides from 4-mers to 52-mers Glycobiology. 2002; 12 (7): 421-6), and the concentration was adjusted by physiological saline. In addition, physiological saline was prepared as a control.

In this example, the test substance or physiological saline was given orally to mice that developed osteoarthritis once a day with an oral probe for 2 weeks.

Macroscopic examination.

The effect in the treatment of osteoarthritis was determined macroscopically. More specifically, Chinese mascara was applied to the surface of the cartilage of the knee joint (tibia and femur) of all animals of each group and got wet with gauze. Chinese mascara remaining in the study area was studied macroscopically using the blind method and animals were classified into 5 categories (table 1). For a description of this assessment method, see Chang DG, Iverson EP, Schinagl RM, Sonoda M, Amiel D, Coutts RD, Sah RL. Quantitation and localization of cartilage degeneration following the induction of osteoarthritis in the rabbit knee. Osteoarthritis Cartilage. 1997 Sep; 5 (5): 357-72.

Table 1 SCORES PER CENT OF REMAINING CHINESE CARCASES one 0-20% 2 20-40% 3 40-60% four 60-80% 5 80-100%

Results.

The results of a macroscopic examination of osteoarthritis are shown in table 2.

table 2 Points for each animal Group receiving physiological saline 5, 5, 4, 5, 5 Average score 4, 8 The group receiving HA4 1, 2, 1, 2, 1 Average score 1, 4

Based on this example, the therapeutic effect in the treatment of osteoarthritis in the HA4 group was confirmed. This result has demonstrated that a drug containing hyaluronan as an active ingredient suppresses osteoarthritis.

Claims (6)

1. A therapeutic agent for the treatment of osteoarthritis containing hyaluronan, comprising from 2 to 20 sugar units as an active ingredient. 2. The therapeutic agent for osteoarthritis according to claim 1, wherein the hyaluronan is a tetrasaccharide containing two units with a single unit in the form of: -D-glucuronic acid-1,3-D-N-acetylglucosamine-1,4-. 3. A therapeutic agent for treating osteoarthritis according to claim 1, which is an oral dosage form. 4. A method of treating osteoarthritis, comprising the step of administering an effective amount of hyaluronan, comprising from 2 to 20 sugar units, to an object in need of such treatment. 5. The method according to claim 4, in which the hyaluronan is a tetrasaccharide comprising two units with a single unit in the form of: -D-glucuronic acid-in-1,3-D-N-acetylglucosamine-in-1,4-. 6. The method according to claim 4, in which hyaluronan is administered orally.