RU2669565C2 - Extended-release formulation for reducing the frequency of urination and method of use thereof - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions relates to medicine, namely to urology, and can be used to reduce the frequency of urination. For this, a pharmaceutical composition comprising effective amounts of zolpidem formulated for immediate release, and one or more analgesic agents formulated for extended or delayed release. Also provided is a pharmaceutical composition consisting of effective amounts of zolpidem formulated for immediate release, and one or more analgesic agents formulated for extended or delayed release.

EFFECT: group of inventions provides the effective suppression of smooth muscle contraction of the bladder.

2 cl, 9 tbl, 8 ex, 1 dwg

Description

Priority is claimed for this application for US Patent Application Registration No. 13/424000, filed March 19, 2012, currently U.S. Patent No. 8236857, and US Patent Application, Registration No. 13/487348, filed June 4, 2012. Full Content the aforementioned applications are incorporated herein by reference.

FIELD OF TECHNOLOGY

The present application, in General, relates to methods and compositions for suppressing muscle contraction and, in particular, to methods and compositions for suppressing smooth muscle contraction of the bladder.

BACKGROUND

The detrusor muscle is a layer of the wall of the bladder, consisting of smooth muscle fibers collected in spiral, longitudinal and circular bundles. When the bladder is stretched, a signal is sent to the parasympathetic nervous system to contract the detrusor muscle. This stimulates the removal of urine from the bladder through the urethra.

For urine to flow out of the bladder, both an autonomously controlled internal sphincter and a consciously controlled external sphincter must be open. Problems with these muscles can lead to urinary incontinence. If the amount of urine reaches 100% of the absolute capacity of the bladder, a consciously controlled sphincter becomes uncontrollable and urine immediately flows out.

An adult bladder typically holds approximately 300-350 ml of urine (working volume), but a full adult bladder can hold up to about 1000 ml of urine (absolute volume), with individual variations. As urine accumulates, the crests resulting from folding the bladder wall (folds) flatten and the bladder wall becomes thinner when stretched, which allows the bladder to hold large amounts of urine without significantly increasing internal pressure.

In most people, the urge to urinate usually occurs when the volume of urine in the bladder reaches approximately 200 ml. At this stage, it is not difficult for a person, if necessary, to resist the urge to urinate. As the bladder continues to fill up, the urge to urinate becomes stronger and harder to ignore. Ultimately, the bladder becomes full to the point where the urge to urinate becomes insurmountable and the person is no longer able to ignore them. Some people want to urinate when the bladder is less than 100% full. Such an increased desire to urinate can interfere with normal activities, including the ability to sleep for quite long periods of uninterrupted rest. In some cases, such an increased desire to urinate may be due to medical conditions, such as benign prostatic hyperplasia or prostate cancer in men, or pregnancy in women. However, an increased desire to urinate also occurs in people, both men and women, who do not suffer from another medical condition.

Accordingly, there is a need for compositions and methods for treating men and women suffering from urge to urinate when their bladder is less than 100% full. These compositions and methods are needed to suppress muscle contraction so that urination may occur in these people when the volume of urine in the bladder exceeds about 100% of its working volume.

SUMMARY OF THE INVENTION

One aspect of the present application relates to a method for decreasing the frequency of urination. The method comprises administering to a subject in need of a pharmaceutical composition comprising: an active ingredient comprising one or more analgesic agents in an amount of 50-400 mg for each agent, said one or more analgesic agents being selected from the group consisting of aspirin, ibuprofen, naproxen, naproxen sodium, indomethacin, nabumetone and acetaminophen, wherein said pharmaceutical composition is formulated for sustained release, so that said active ingredient is released on continuously for 5-24 hours. The method can be used to treat nocturia or an overactive bladder.

Another aspect of the present application relates to a method for decreasing the frequency of urination. The method comprises administering to a subject in need of a pharmaceutical composition comprising: an active ingredient comprising one or more analgesic agents in an amount of 50-400 mg for each agent, said one or more analgesic agents being selected from the group consisting of aspirin, ibuprofen, naproxen, naproxen sodium, indomethacin, nabumetone and acetaminophen, wherein said pharmaceutical composition is formulated for sustained release, characterized by a biphasic release profile, in which 20-60% of the specified active ingredient is released within two hours after administration and the remainder of the specified active ingredient is released continuously for 5-24 hours. The method can be used to treat nocturia or an overactive bladder.

Another aspect of the present application relates to a method for decreasing the frequency of urination, comprising administering to a subject in need of an effective amount of botulinum toxin, said botulinum toxin being injected into a muscle of the bladder, and orally administering to said subject a pharmaceutical composition comprising: an active ingredient comprising one or more analgesic agents in an amount of 50-400 mg for each agent, wherein said one or more analgesic agents are selected from the group consisting of ina, ibuprofen, naproxen, naproxen sodium, indomethacin, nabumetone and acetaminophen, wherein said pharmaceutical composition is formulated for sustained release. The method can be used to treat nocturia or an overactive bladder.

Another aspect of the present application relates to a method for decreasing the frequency of urination, comprising administering to a subject in need thereof an effective amount of one or more analgesic agents and an effective amount of zolpidem. The method can be used to treat nocturia or an overactive bladder.

Another aspect of the present application relates to a method for decreasing the frequency of urination, comprising administering to a subject a pharmaceutical composition comprising: one or more analgesic agents and an antidiuretic, wherein said one or more analgesic agents are formulated for delayed release and said antidiuretic is formulated for immediate release. The method can be used to treat nocturia or an overactive bladder.

Another aspect of the present application relates to a pharmaceutical composition comprising an active ingredient comprising one or more analgesic agents, zolpidem and a pharmaceutically acceptable carrier.

Another aspect of the present application relates to a pharmaceutical composition comprising one or more analgesic agents and an antidiuretic, wherein said one or more analgesic agents are formulated for delayed release and said antidiuretic is formulated for immediate release.

BRIEF DESCRIPTION OF THE DRAWINGS

In FIG. 1A and 1B are diagrams showing that analgesics regulate the expression of costimulatory molecules by Raw 264 macrophage cells in the absence (Fig. 1A) or in the presence (Fig. 1B) of LPS. Cells were cultured for 24 hours in the presence of only an analgesic or analgesic together with LPS Salmonella typhimurium (0.05 μg / ml). The results are average relative values in% for CD40 + CD80 + cells.

DETAILED DESCRIPTION OF THE INVENTION

The following detailed description is provided in order to help those skilled in the art to implement and apply the present invention. For purposes of explanation, specific nomenclature is provided to provide a thorough understanding of the present invention. However, it will be apparent to those skilled in the art that these specific details are not necessary for the practice of the invention. Descriptions of specific applications are provided only as illustrative examples. The present invention should not be limited by the above embodiments, but should correspond to the broadest possible scope, consistent with the principles and features disclosed herein.

As used herein, the term “effective amount” means the amount necessary to achieve a selected result.

As used herein, the term “analgesic” refers to agents, compounds, or drugs used to alleviate pain, including anti-inflammatory compounds. Typical analgesic and / or anti-inflammatory drugs, compounds, or drugs include, but are not limited to, the following substances: non-steroidal anti-inflammatory drugs (NSAIDs), salicylates, aspirin, salicylic acid, methyl salicylate, diflunisal, salsalate, olsalazine, sulfasalazoline acetanilide, acetaminophen, phenacetin, phenamates, mefenamic acid, meclofenamate, sodium meclofenamate, heteroaryl acetic acid derivatives, tolmetin, ketorolac, diclofenac, pro zvodnye propionic acid, ibuprofen, naproxen sodium, naproxen, fenoprofen, ketoprofen, flurbiprofen, oxaprozin; enolic acids, derivatives of oxicam, piroxicam, meloxicam, tenoxicam, ampiroxicam, droxikam, pivoxicam, pyrazolone derivatives, phenylbutazone, oxyphenbutazone, antipyrine, aminopyrine, dipyrone, coxibutin, sucecibenopropylbenzene, isofecinoptene, nofecacinoptene, nofecacinoptene, nrofecacinoptene, nofecacinoptene, nofecacinoptene lumiracoxib, etoricoxib, parecoxib, valdecoxib, tiracoxib, etodolac, darbufelon, dexketoprofen, aceclofenac, lycofelon, bromfenac, loxoprofen, pranoprofen, piroxicam, nimesulide, cisolirtine, 3-formylamino nilamino-6-phenoxy-4H-1-benzopyran-4-one, meloxicam, lornoxicam, d-indobufen, mofesolac, amtolmethine, pranoprofen, tolfenamic acid, flurbiprofen, suprofen, oxaprozin, zaltoprofen, aluminofrofen, tylaminofrofen, tylaminofrofen, tylaminofrofen, tylaminofrofen, tylaminofrofen, tymnofenofen, tylaminofrofen, tylaminofrofen, tylaminofrofen, tylaminofrofen, tymnofenofen, tymnofenofen, tymnofenofen, tymnofenofen, tymnofenofen, tylaminofrofen, tymnoprofen its hydrates and its solvates.

As used herein, the terms “coxib” and “COX inhibitor” refer to a composition of compounds that are capable of inhibiting the activity or expression of COX2 enzymes or are capable of inhibiting or reducing the severity, including pain and edema, of a severe inflammatory response.

As used herein, the term “derivative” refers to a chemically modified compound in which the modification is recognized by a conventional qualified chemist, for example, an ester or acid amide, protecting groups, for example, a benzyl group for an alcohol or thiol and a tert-butoxycarbonyl group for an amine.

As used herein, the term “analogue” refers to a compound that is a chemically modified form of a particular compound or class thereof and which retains pharmaceutical and / or pharmacological activities characteristic of the specified compound or class.

As used herein, the term “pharmaceutically acceptable salts” refers to derivatives of the described compounds in cases where the parent compound is modified by the formation of acidic or basic salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, salts of mineral or organic acids with bases such as amines; salts of alkalis or organic bases with acids such as carboxylic acids, and the like. Pharmaceutically acceptable salts include conventional non-toxic salts or quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include salts derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and salts derived from organic acids, such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilibenzoic, 2- fumaric, toluenesulfonic, methanesulfonic, ethanedisulfonic, oxalic, isethionic and the like.

As used herein, the term “pharmaceutically acceptable” is used with reference to compounds, materials, compositions and / or dosage forms which, in the framework of sound medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions or other problems or complications, with a reasonable benefit / risk ratio.

As used herein, the term “subject” or “patient” encompasses mammals. In one aspect, the mammal is human. In another aspect, the mammal is a non-human primate, such as chimpanzees and other apes and apes. In one aspect, the mammal is a domestic animal, such as a rabbit, dog, or cat. In another aspect, the mammal is a farm animal, such as cattle, horse, sheep, goat or pig. In another aspect, the mammal is a laboratory animal, including rodents such as rats, mice, and guinea pigs, and the like.

The bladder has two important functions: storage of urine and emptying. Storage of urine occurs at low pressure, which means that the detrusor muscle relaxes during the filling phase. Bladder emptying requires a coordinated detrusor contraction and relaxation of the urethral sphincter muscles. Impairment of storage function can lead to symptoms of the lower urinary tract, such as urgent urination, frequent urination and urinary incontinence, components of the syndrome of overactive bladder. Overactive bladder syndrome, which may be associated with involuntary contractions of the smooth muscles of the bladder (detrusor) during storage, is a common and silent problem, the prevalence of which has only recently been estimated.

One aspect of the present application relates to a method for decreasing the frequency of urination by administering to a person in need thereof a pharmaceutical composition formulated in a sustained release formulation. The pharmaceutical composition contains one or more analgesic agents and optionally one or more antimuscarinic agents, one or more antidiuretic agents, one or more antispasmodics and / or zolpidem. The method can be used to treat nocturia or an overactive bladder.

"Sustained Release", also known as Slow Release (SR), Slow Release (SA), Timed Release (TR), Controlled Release (CR), Modified Release (MR) or Continuous Release (CR), is used in medicine is a mechanism where tablets or capsules dissolve slowly and release the active ingredient over time. The benefits of sustained release tablets or capsules are that they can often be taken less frequently than immediate release drugs of the same drug, and that they maintain more stable levels of the drug in the blood, thereby prolonging the duration of the drug and lowering the peak amount of the drug in the blood. For example, a long-release analgesic may allow a person to sleep all night without waking up to use the toilet.

In one embodiment, the pharmaceutical composition is formulated for sustained release by placing the active ingredient in a matrix of insoluble matter (s), such as acrylics or chitin. The sustained release form is intended to release the analgesic compound at a given rate in order to maintain a constant level of the drug for a certain period of time. This can be achieved using various drugs, including, but not limited to, liposomes and drug-polymer conjugates, such as hydrogels.

A sustained release preparation can be designed to release the active agents at a given rate so as to maintain a constant level of the drug for a certain, long period of time, for example, up to about 24 hours, about 20 hours, about 16 hours, about 12 hours, about 10 hours, about 9 hours, about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours or about 1 hour after administration or after the lag period, St. delayed-release drug.

In some preferred embodiments, the active agents are released over a period of time from about 2 to about 10 hours. Alternatively, the active agents may be released over about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10 hours, about 12 hours, about 16 hours, about 20 hours, or about 24 hours. In other embodiments, active agents are released over a period of about three to about eight hours after administration.

In some embodiments, the sustained release preparation comprises an active core consisting of one or more inert particles, each in the form of a ball, granule, pill, granular particle, microcapsule, microsphere, microgranule, nanocapsule or nanosphere, with the drug applied to its surface in the form , for example, a drug-containing coating or film-forming composition, using, for example, a fluidized bed technique or other methodologies known to those skilled in the art oh area. An inert particle can have a different size, provided that it is large enough to remain poorly soluble. Alternatively, the active core can be obtained by granulation and grinding and / or by extrusion and spheronization of a polymer composition containing a drug.

Active agents can be introduced into an inert carrier using methods known to one of skill in the art, such as drug layering, powder coating, extrusion / spheronization, rolling or granulation. The amount of drug in the core will depend on the dose that is required, and usually varies from about 5 to 90 mass%. Typically, the polymer coating on the active core will be from about 1 to 50%, based on the weight of the coated particles, depending on the required lag period and / or selected polymers and solvents for the coating. Specialists in this field will be able to choose the appropriate amount of drug for coating or inclusion in the core to achieve the desired dosage. In one embodiment, the inactive core may be a sugar sphere or buffer crystal, or an encapsulated buffer crystal, for example, calcium carbonate, sodium bicarbonate, fumaric acid, tartaric acid, and so on, which changes the microenvironment of the drug to facilitate its release.

In sustained release formulations, various coatings or sustained release mechanisms can be used to promote the gradual release of the active agents over time. In some embodiments, the sustained release agent comprises a polymer that controls release by controlled dissolution. In a specific embodiment, the active agent (a) is included in a matrix containing an insoluble polymer and particles or granules of the drug coated with polymeric materials of various thicknesses. The polymeric material may contain a lipid barrier including a wax material such as carnauba wax, beeswax, spermaceti wax, candelilla wax, shellac wax, cocoa butter, cetostearyl alcohol, partially hydrogenated vegetable oils, ceresin, paraffin wax, ceresin, myristyl alcohol, stear alcohol, cetyl alcohol and stearic acid, along with surfactants such as polyoxyethylene sorbitan monooleate. Upon contact with an aqueous medium, such as biological fluids, the polymer coating emulsifies or erodes after a predetermined lag period, depending on the thickness of the polymer coating. The lag period is independent of gastrointestinal motility, pH, or being in the stomach.

In other embodiments, the sustained release agent comprises a polymer matrix that provides controlled release through diffusion. The matrix may contain one or more hydrophilic and / or water-swellable matrix-forming polymers, pH-dependent polymers and / or pH-independent polymers.

In one embodiment, the sustained release preparation comprises a water-soluble or water-swellable, matrix forming polymer, optionally containing one or more solubility enhancing excipients and / or release agents. Upon solubilization of the water-soluble polymer, the active agent (a) dissolves (if it is soluble) and gradually diffuses through the hydrated part of the matrix. The gel layer increases over time as more and more water penetrates the core of the matrix, increasing the thickness of the gel layer and providing a diffusion barrier for the release of the drug. When the outer layer becomes fully hydrated, the polymer chains become completely relaxed and can no longer maintain the integrity of the gel layer, which leads to the dissolution and erosion of the outer hydrated polymer on the surface of the matrix. Water continues to penetrate the core through the gel layer until it completely erodes. While soluble drugs are released through this combination of diffusion and erosion mechanisms, erosion is the predominant mechanism for insoluble drugs, regardless of dose.

Similarly, water-swellable polymers tend to hydrate and swell in body fluids, forming a homogeneous matrix structure that retains its shape during drug release and serves as a carrier for the drug, solubility enhancers and / or release stimulants. The result of the initial phase of hydration of the matrix polymer is a slow release of the drug (lag phase). After the water-swellable polymer becomes fully hydrated and swollen, the water inside the matrix can similarly dissolve the drug and make it possible to diffuse outward through the matrix coating.

In addition, the porosity of the matrix can be increased by flushing out pH-dependent release stimulators so that the drug is released at a faster rate. Then, the drug release rate becomes constant and depends on the diffusion of the drug through the hydrated polymer gel. The rate of release from the matrix depends on various factors, including the type and level of polymer content; solubility and dose of the drug; polymer: drug ratio; type and level of filler content; the ratio of polymer to filler; particle size of drug and polymer, as well as porosity and matrix shape.

Typical hydrophilic and / or water-swellable matrix-forming polymers include, but are not limited to, cellulosic polymers, including hydroxyalkyl celluloses and carboxyalkyl celluloses, such as hydroxypropyl methyl cellulose (HPMC), hydroxypropyl cellulose (HPC), hydroxyethyl cellulose (HEC), methyl cellulose (carboxyl cellulose (HEC), methyl cellulose (cellulose) CMC), powdered cellulose, such as microcrystalline cellulose, cellulose acetate, ethyl cellulose, its salts, and combinations with it; alginates, gums, including heteropolysaccharide gums and homopolysaccharide gums, such as xanthan gum, tragacanth, pectin, gum arabic, karaya gum, alginates, agar, guar gum, hydroxypropyl guar, wigum, carrageenan, gum, fruit gum, fruit gum them; acrylic resins, including polymers and copolymers of acrylic acid, methacrylic acid, methyl acrylate and methyl methacrylate, as well as crosslinked derivatives of polyacrylic acid, such as carbomers (e.g. Carbopol ^® , including Carbopol ^® 71G NF, various grades depending on molecular weight are supplied by Noveon, Inc., Cincinnati, Ohio); carrageenan; polyvinyl acetate (e.g., KOLLIDON ^® SR); polyvinylpyrrolidone and its derivatives, such as crospovidone; polyethylene oxides and polyvinyl alcohol. Preferred hydrophilic and water-swellable polymers include cellulosic polymers, especially HPMC.

The sustained release preparation may further comprise at least one binder that is capable of crosslinking the hydrophilic compound to form a hydrophilic polymer matrix (i.e., a gel matrix) in an aqueous medium, including biological fluids.

Typical binders include homopolysaccharides such as galactomannan gums, guar gum, hydroxypropyl guar gum, hydroxypropyl cellulose (HPC; for example, Klucel EXF) and locust bean gum. In other embodiments, the binder is an alginic acid derivative, HPC, or microcrystalline cellulose (MCC). Other binders include, but are not limited to, starches, microcrystalline cellulose, hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose and polyvinylpyrrolidone.

In one embodiment, the incorporation method is layering a drug by spraying a suspension of the active agent (s) and a binder onto an inert carrier.

The binder may be present in the granular preparation in an amount of from about 0.1% to about 15% by weight and preferably from about 0.2% to about 10% by weight.

In some embodiments, the hydrophilic polymer matrix may further comprise an ionic polymer, a non-ionic polymer, or a water-insoluble hydrophobic polymer to create a stronger gel layer and / or to reduce the number and size of pores in the matrix in order to reduce diffusion and erosion and concomitant release rates active funds (funds). In addition, this can suppress the initial effect of the explosion and create a more stable “zero order release” of the active agent (s).

Typical ionic polymers to reduce the dissolution rate include both anionic and cationic polymers. Representative anionic polymers include, e.g., sodium carboxymethylcellulose (Na CMC), sodium alginate, acrylic acid polymers or carbomers (e.g., Carbopol ^® 934, 940, 974P NF); enteric polymers such as polyvinyl acetate phthalate (PVAP), methacrylic acid copolymers (e.g., EUDRAGIT ^® L100, L 30D 55, A and FS 30D), hypromellose acetate succinate (AQUAT HPMCAS) and xanthan gum. Typical cationic polymers include, for example, dimethylaminoethyl methacrylate copolymer (e.g., EUDRAGIT ^® E 100). The inclusion of anionic polymers, especially enteric polymers, offers the advantage of creating a pH independent release profile for slightly alkaline drugs compared to a hydrophilic polymer alone.

Typical non-ionic polymers for slowing the dissolution rate include, for example, hydroxypropyl cellulose (HPC) and polyethylene oxide (PEO) (e.g. POLYOX ™).

Exemplary hydrophobic polymers include ethyl cellulose (e.g., ETHOCEL ™, SURELEASE ^®), cellulose acetate, methacrylic acid copolymers (e.g., EUDRAGIT ^® NE 30D), copolymers ammoniometakrilata (e.g., EUDRAGIT ^® RL 100 or PO RS100), polyvinyl acetate, glyceryl monostearate, fatty acid such as acetyl tributyl citrate, as well as combinations and derivatives thereof.

The swellable polymer may be included in the preparation in a proportion of from 1% to 50% by weight, preferably from 5% to 40% by weight, most preferably from 5% to 20% by weight. Swellable polymers and binders can be included in the preparation either before or after granulation. The polymers can also be dispersed in organic solvents or hydroalcohols and sprayed during the granulation process.

Typical release-promoting agents comprising pH-dependent enteric polymers that remain intact at pHs lower than about 4.0 and dissolve at pHs higher than 4.0, preferably higher than 5.0, most preferably about 6 , 0, are considered useful as release-promoting agents for the present invention. Exemplary pH-sensitive polymers include, but are not limited to, copolymers of methacrylic acid copolymers, methyl methacrylate-methacrylic acid (e.g., EUDRAGIT ^® L100 (type A), EUDRAGIT ^® S100 (type B), Rohm GmbH, Germany); copolymers of methacrylic acid-ethyl acrylate copolymer (e.g., EUDRAGIT ^® L100-55 (type C) and EUDRAGIT ^® L30D-55 dispersion copolymers, Rohm GmbH, Germany); copolymers of methacrylic acid-methyl methacrylate and methyl methacrylate (EUDRAGIT ^® FS); terpolymers of methacrylic acid, methacrylate and ethyl acrylate; cellulose acetate phthalates (CAP); hydroxypropyl methylcellulose phthalate (HPMCP) (e.g., HP-55, HP-50, HP-55S, Shinetsu Chemical, Japan); polyvinyl acetate phthalate (PVAP) (e.g., COATERIC ^®, OPADRY ^® enteric White OY-P-7171); polyvinyl butyrate acetate; cellulose acetate succinates (CAS); hydroxypropylmethylcellulose acetate succinate (HPMCAS), e.g., HPMCAS category LF, MF category, HF categories including AQOAT ^® LF and ^{AQOAT ® MF (Shin-Etsu Chemical} , Japan); shellac (e.g. MARCOAT ™ 125 and MARCOAT ™ 125N); vinyl acetate-maleic anhydride copolymer; styrene-maleic monoester copolymer; carboxymethyl ethyl cellulose (CMEC, Freund Corporation, Japan); cellulose acetate phthalates (CAP) (e.g. AQUATERIC ^® ); cellulose acetate trimellitates (CAT); as well as mixtures of two or more of them in mass ratios from about 2: 1 to about 5: 1, such as, for example, a mixture of EUDRAGIT ^® L 100-55 and EUDRAGIT ^® S 100 in a mass ratio from about 3: 1 to about 2: 1 or a mixture of EUDRAGIT ^® L 30 D-55 and EUDRAGIT ^® FS in a weight ratio of from about 3: 1 to about 5: 1.

These polymers can be used either individually or in combination or together with polymers other than those mentioned above. Preferred enteric pH dependent polymers are pharmaceutically acceptable methacrylic acid copolymers. These copolymers are anionic polymers based on methacrylic acid and methyl methacrylate and preferably have an average molecular weight of about 135,000. The ratio of free carboxyl groups to methyl esterified carboxyl groups in these copolymers can range from, for example, from 1: 1 to 1: 3, for example , approximately 1: 1 or 1: 2. Such polymers are sold under the trade name EUDRAGIT ^® , e.g. EUDRAGIT ^® L-series, e.g. EUDRAGIT L 12.5 ^® , EUDRAGIT L 12.5P ^® , EUDRAGIT L100 ^® , EUDRAGIT L 100-55 ^® , EUDRAGIT L-30D ^® , EUDRAGIT L-30 D-55 ^® , EUDRAGIT S ^® series, e.g. EUDRAGIT S 12.5 ^® , EUDRAGIT S 12.5P ^® , EUDRAGIT S100 ^® . Release promoters are not limited to pH dependent polymers. Other hydrophilic molecules that quickly dissolve and quickly wash out of the dosage form, leaving a porous structure, can also be used for the same purpose.

In some embodiments, the matrix may comprise a combination of release promoters and solubility enhancers. Solubility enhancers can be ionic and nonionic surfactants, complexing agents, hydrophilic polymers, pH modifiers, such as acidifying agents and alkalizing agents, as well as molecules that increase the solubility of poorly soluble drugs due to molecular encapsulation. Several solubility enhancers can be used at the same time.

Solubility enhancers may include surfactants such as sodium docusate, sodium lauryl sulfate, sodium stearyl fumarate, twins ^® and spans (PEO-modified sorbitan monoesters and sorbitan fatty acid esters), poly (ethylene oxide) -polypropylene oxide-poly (ethylene) block copolymers ) (also called pluronics ™); complexing agents such as low molecular weight polyvinyl pyrrolidone and low molecular weight hydroxypropyl methyl cellulose; molecules that promote solubility through molecular encapsulation, such as cyclodextrins, and pH modifying agents, including acidifying agents, such as citric acid, fumaric acid, tartaric acid, and hydrochloric acid; and alkalizing agents such as meglumine and sodium hydroxide.

The solubility enhancers are typically from 1% to 80% by weight, preferably from 1% to 60%, more preferably from 1% to 50% of the dosage form and can be incorporated in various ways. They can be included in the preparation before granulation in dry or wet form. They can also be added to the preparation after the remaining materials are granulated or otherwise processed. During the granulation process, the solubilizers can be sprayed in the form of solutions with or without the addition of a binder.

In one embodiment, the sustained release preparation comprises, on top of the active core, a water-insoluble water-permeable polymer coating or matrix containing one or more water-insoluble water-permeable film-forming substances. In addition, the coating may contain one or more water-soluble polymers and / or one or more plasticizers. The water-insoluble polymer coating is a barrier coating for the release of active agents in the core, while the category of substances with a low molecular weight (viscosity) has higher release rates compared to the category of substances with a higher viscosity.

In preferred embodiments, the water insoluble film forming polymers include one or more esters of an alkylcellulose, e.g., ethylcellulose, and mixtures thereof (e.g., ethylcellulose categories PR100, PR45, PR20, PR10 and PR7; ETHOCEL ^®, Dow).

Typical water soluble polymers include polyvinylpyrrolidone (POVIDONE ^®), hydroxypropylmethylcellulose, hydroxypropylcellulose and mixtures thereof.

In some embodiments, the water-insoluble polymer imparts suitable properties (e.g., sustained release characteristics, mechanical properties, and coating properties) without the need for a plasticizer. For example, coatings containing polyvinyl acetate (PVA), neutral acrylate / methacrylate ester copolymers such as the commercially available EUDRAGIT NE30D from Evonik Industries, ethyl cellulose in combination with hydroxypropyl cellulose, waxes and the like, can be used without plasticizers.

In another embodiment, the water-insoluble polymer matrix may further comprise a plasticizer. The amount of plasticizer required depends on the plasticizer, the properties of the water-insoluble polymer and the desired final coating properties. Suitable levels of plasticizer are in the range of from about 1% to about 20%, from about 3% to about 20%, from about 3% to about 5%, from about 7% to about 10%, from about 12% to about 15 %, from about 17% to about 20% or about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10% , about 15% or about 20% by weight relative to the total weight of the coating, including all ranges and subranges between the indicated values.

Typical plasticizers include, but are not limited to, triacetin, acetylated monoglyceride, oils (castor oil, hydrogenated castor oil, rapeseed oil, sesame oil, olive oil and so on), citrate esters, triethyl citrate, acetyl triethyl citrate, acetyl citrate, trityl citrate n-butyl citrate, diethyl phthalate, dibutyl phthalate, dioctyl phthalate, methyl paraben, propyl paraben, propyl paraben, butyl paraben, diethyl sebacate, dibutyl sebacate, glycerol tributyrate, substituted triglycerides and glycerol monoacetylated and diacetylated glycerides (e.g. MYVACET ^® 9-45), glyceryl monostearate, glycerol tributyrate, polysorbate 80, polyethylene glycol (such as PEG-4000, PEG-400), propylene glycol, 1,2-propylene glycol, glycerol, sorbitol diethyl malate, diethyl fumarate, diethyl malonate, dibutyl succinate, fatty acids, glycerin, sorbitol, diethyl oxalate, diethyl malate, diethyl maleate, diethyl fumarate, diethyl succinate, diethyl malonate, dioctyl phthalate, and mixtures thereof, dibutyl The plasticizer may have surfactant properties, so that it can act as a release modifier. For example, non-ionic detergents such as Brij 58 (polyoxyethylene (20) cetyl ether) and the like can be used.

Plasticizers can be high boiling point organic solvents that are used to give flexibility to hard or brittle polymeric materials and which can affect the release profile of the active agent (s). Plasticizers typically cause weakening of the intermolecular bonding forces along the polymer chains, which leads to various changes in the properties of the polymers, including a decrease in tensile strength, an increase in tension, and a decrease in the glass transition temperature or softening temperature of the polymer. The amount and choice of plasticizer may affect the strength of the tablet, for example, and may even affect its dissolution or decomposition characteristics, as well as its physical and chemical stability. Some plasticizers can increase the elasticity and / or flexibility of the coating, thereby reducing the brittleness of the coating.

In another embodiment, the sustained release preparation comprises a combination of at least two gelling polymers, including at least one non-ionic gelling polymer and / or at least one anionic gelling polymer. A gel formed by a combination of gelling polymers provides controlled release such that when the drug is swallowed and comes into contact with gastrointestinal fluids, the polymers closest to the surface hydrate to form a viscous gel layer. Due to the high viscosity, the viscous layer dissolves only gradually, causing the same process in the material underneath. Thus, the mass dissolves slowly, thereby slowly releasing the active ingredient into the gastrointestinal fluids. The combination of at least two gelling polymers allows you to manipulate the properties of the resulting gel, such as viscosity, to achieve the desired release profile.

In a specific embodiment, the preparation comprises at least one non-ionic gelling polymer and at least one anionic gelling polymer. In another embodiment, the preparation contains two different non-ionic gelling polymers. In another embodiment, the preparation contains a combination of nonionic gelling polymers of the same chemical composition, but having different solubilities, viscosities and / or molecular weights (e.g., a combination of hydroxypropyl methylcelluloses of different viscosity grades, e.g., HPMC K100 and HPMC K15M or HPMC K100M).

Typical anionic gelling polymers include, but are not limited to, sodium carboxymethyl cellulose (Na CMC), carboxymethyl cellulose (CMC), anionic polysaccharides such as sodium alginate, alginic acid, pectin, polyglucuronic acid (poly-α- and β-1, 4-glucuronic acid), polygalacturonic acid (pectinic acid), chondroitin sulfate, carrageenan, furcellaran, anionic gums such as xanthan gum, acrylic acid polymers or carbomers (Carbopol ^® 934, 940, 974P NF), copolymers Carbopol ^®, Pemulen polymer ^® , polycarbophil and others.

Typical non-ionic gelling polymers include, but are not limited to, povidone (PVP: polyvinyl pyrrolidone), polyvinyl alcohol, a copolymer of PVP and polyvinyl acetate, HPC (hydroxypropyl cellulose), HPMC (hydroxypropyl methyl cellulose), hydroxyethyl cellulose, hydroxymethyl gum, hydroxymethyl gum, hydroxymethyl gum, hydroxymethyl gum, hydroxymethyl gum, hydroxymethyl gum, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, polyhydroxyethyl methacrylate (PHEMA), water-soluble non-ionic polymethacrylates and their copolymers, modified cellulose, modified polysaccharides, non-ionic gums, non-ionic polysaccharides and / or mixtures thereof and.

The preparation may optionally contain an enteric polymer as described above and / or at least one excipient, such as a filler, a binder (described above), a disintegrating agent and / or a flow agent or a glidant.

Typical excipients include, but are not limited to, lactose, glucose, fructose, sucrose, dicalcium phosphate, sugar alcohols, also known as “sugar polyols,” such as sorbitol, mannitol, lactitol, xylitol, isomalt, erythritol, and hydrogenated starch hydrolysates ( a mixture of several sugar alcohols), corn starch, potato starch, sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate, enteric polymers, or mixtures thereof.

Typical binders include, but are not limited to, water-soluble hydrophilic polymers such as povidone (PVP: polyvinylpyrrolidone), copovidone (a copolymer of polyvinylpyrrolidone and polyvinyl acetate), low molecular weight HPC (hydroxypropyl cellulose, low molecular weight cellulose hydroxymethyl cellulose cellulose methylcellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose methyl cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose cellulose polyethylene oxide, gum arabic, dextrin, magnesium aluminum silicate, starch and polymethacrylates such as EUDRAGIT NE 30D, EUDRAGIT RL, EUDRAGIT RS, EUDRAGIT E, polyvinyl acetate and enteric polymers, or mixtures thereof.

Typical disintegrants include, but are not limited to, low-substituted sodium carboxymethyl cellulose, crospovidone (crosslinked polyvinylpyrrolidone), sodium carboxymethyl starch (sodium starch glycolate, 1500 starch sodium, carboxymethyl cellulose cellulose) in water, starch, calcium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, as well as magnesium or aluminum silicate.

Typical glidants include, but are not limited to, magnesium, silica, talc, starch, titanium dioxide, and the like.

In yet another embodiment, a sustained release preparation is prepared by coating particles containing a water-soluble / dispersible drug, such as a granule or a combination of granules (as described above), a coating material and optionally a blowing agent and other excipients. The coating material is preferably selected from the group consisting of cellulosic polymers such as ethyl cellulose (e.g., SURELEASE ^®), methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, cellulose acetate and cellulose acetate phthalate; polyvinyl alcohol; acrylic polymers such as polyacrylates, polymethacrylates and their copolymers, as well as other coating materials based on water or based on a solvent. The release control coating for a particular population of granules can be controlled by at least one parameter of the release control coating, for example, the nature of the coating, level of coating, type and concentration of pore former, process parameters, and combinations thereof. Thus, a change in a parameter, such as the concentration of a blowing agent or curing conditions, allows you to make changes in the release of the active agent (s) from any particular set of granules, thereby making it possible to selectively adjust the drug to achieve a given release profile.

Pore formers suitable for use in the release control coating described herein may be organic or inorganic substances and include materials that can dissolve, extract, or wash out of the coating in an application environment. Typical blowing agents include, but are not limited to, organic compounds such as mono-, oligo- and polysaccharides, including sucrose, glucose, fructose, mannitol, mannose, galactose, sorbitol, pullulan, dextran; environmentally soluble polymers such as water-soluble hydrophilic polymers, hydroxyalkyl celluloses, carboxyalkyl celluloses, hydroxypropyl methyl cellulose, cellulose ethers, acrylic resins, polyvinyl pyrrolidone, crosslinked polyvinyl pyrrolidone, polyethylene oxide, carboxylic polyols, polyethylene polyglycol, polyol polyglycol, polypropylene glycols or their block polymers, polyglycols, poly (α-Ω) alkylenediols; inorganic compounds such as alkali metal salts, lithium carbonate, sodium chloride, sodium bromide, potassium chloride, potassium sulfate, potassium phosphate, sodium acetate, sodium citrate, suitable calcium salts, combinations thereof and the like.

The release control coating may further contain other additives known in the art, such as plasticizers, release agents, glidants (or flow agents), and antifoam agents.

In some embodiments, the coated particles or granules may also have an “outer coating” to provide, for example, moisture protection, reduce static charge, mask taste, aromatize, color and / or polish, or impart other cosmetic attractiveness to the granules. Suitable coating materials for such an external coating are known in the art and include, but are not limited to, cellulosic polymers such as hydroxypropyl methyl cellulose, hydroxypropyl cellulose and microcrystalline cellulose, or combinations thereof (e.g., various OPADRY ^® coating materials).

Coated particles or granules may additionally contain enhancers, for example, but not limited to, solubility enhancers, dissolution enhancers, absorption enhancers, permeability enhancers, stabilizers, complexing agents, enzyme inhibitors, p-glycoprotein inhibitors and multidrug resistance protein inhibitors . Alternatively, the preparation may also contain enhancers that are separated from the coated particles, for example, in a separate collection of granules or in powder form. In another embodiment, the enhancer (s) may be contained in a separate layer on the coated particles either below or above the release control coating.

In other embodiments, a sustained release preparation is formulated to release an active agent (s) using an osmosis mechanism. As an example, a capsule may be formulated with one osmotic unit, or it may include 2, 3, 4, 5, or 6 push-pull units encapsulated in a hard gelatine capsule, whereby each two-layer push-pull unit contains an osmotic push layer and a drug layer surrounded by a semi-permeable the membrane. One or more holes are drilled in the membrane close to the drug layer. This membrane can be further coated with a pH-dependent enteric coating to prevent release until after gastric emptying. The gelatin capsule dissolves immediately after swallowing. When the push-pull unit (s) enters the small intestine, the enteric coating disintegrates, allowing fluid to flow through the semipermeable membrane, while the swellable osmotic ejection compartment pushes drugs through the hole (s) at a speed precisely controlled by the rate of water passing through the semipermeable membrane. Drug release can occur at a constant rate for up to 24 hours or more.

An osmotic ejection layer contains one or more osmotic agents that create a motive force for transporting water through a semipermeable membrane to the core of the drug delivery carrier. One class of osmotic agents includes water-swellable hydrophilic polymers, also called “osmopolymers” and “hydrogels,” including, but not limited to, hydrophilic vinyl and acrylic polymers, polysaccharides such as calcium alginate, polyethylene oxide (PEO), polyethylene glycol (PEG ), polypropylene glycol (PPG), poly (2-hydroxyethyl methacrylate), poly (acrylic) acid, poly (methacrylic) acid, polyvinylpyrrolidone (PVP), crosslinked PVP, polyvinyl alcohol (PVA), PVA / PVP copolymers, PVA / PVP copolymers with hydrophobic monomers such as m ethyl methacrylate and vinyl acetate, hydrophilic polyurethanes containing large blocks of PEO, croscarmellose sodium, carrageenan, hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), hydroxypropyl methyl cellulose (HPMC), carboxymethyl cellulose (CMC, sodium carboxymethyl cellulose, carboxymethyl carboxymethyl carboxymethyl cellulose) xanthan gum and sodium starch glycolate.

Another class of osmotic agents includes osmogens, which are capable of absorbing water, creating an osmotic pressure gradient across a semipermeable membrane. Typical osmogens include, but are not limited to, inorganic salts such as magnesium sulfate, magnesium chloride, calcium chloride, sodium chloride, lithium chloride, potassium sulfate, potassium phosphates, sodium carbonate, sodium sulfite, lithium sulfate, potassium chloride and sodium sulfate; sugars such as dextrose, fructose, glucose, inositol, lactose, maltose, mannitol, raffinose, sorbitol, sucrose, trehalose and xylitol; organic acids such as ascorbic acid, benzoic acid, fumaric acid, citric acid, maleic acid, sebacic acid, sorbic acid, adipic acid, edetic acid, glutamic acid, p-toluenesulfonic acid, succinic acid and tartaric acid; urea, as well as mixtures thereof.

Materials useful for preparing semi-permeable membranes include various categories of acrylics, vinyls, esters, polyamides, polyesters and cellulose derivatives that are water-permeable and insoluble in water at physiological pH values or may become insoluble in water as a result of chemical modification, such as cross stitching.

In some embodiments, a sustained release preparation may contain a polysaccharide coating that is resistant to erosion in both the stomach and intestines. Such polymers can break down only in the colon, which contains a large amount of microflora, containing biodegradation enzymes that destroy, for example, polysaccharide coatings, releasing the drug in a controlled, time-dependent manner. Typical polysaccharide coatings may include, for example, amylose, arabinogalactan, chitosan, chondroitin sulfate, cyclodextrin, dextran, guar gum, pectin, xylan, as well as combinations or derivatives thereof.

In some embodiments, the pharmaceutical composition is formulated for sustained sustained release. As used herein, the term “delayed release” refers to a drug that does not disintegrate and does not release the active ingredient (s) in the body immediately. In some embodiments, the term “sustained sustained release” is used to refer to a drug preparation having a release profile that has a predetermined delayed release of the drug after administration. In some embodiments, the sustained release formulation comprises an enteric-coated sustained release formulation that is a barrier used in the case of oral medications that prevents the release of the medicament until it reaches the small intestine. Delayed release preparations, for example, with an enteric coating, prevent the dissolution of drugs in the stomach that irritate the stomach, such as aspirin. Such coatings are also used to protect drugs that are unstable in an acidic environment from exposure to the acidic environment of the stomach, instead delivering them to an environment with an alkaline pH (in the intestines pH 5.5 and above), where they will not disintegrate and have their desired effect.

The term "pulsed release" refers to one of the types of delayed release, it is used in this document in relation to a drug preparation, which provides a quick and transient drug release for a short period of time immediately after a given lag period, thereby creating a "pulse" plasma drug profile after drug administration. Drugs can be designed to provide a single pulsed release or several pulsed releases at predetermined time intervals after administration, or pulsed release (e.g., 20-60% of the active ingredient) followed by a sustained release over a period of time (e.g., continuous release of the remainder of the active ingredient )

A delayed release or pulsed release preparation typically includes one or more elements coated with a barrier coating that dissolves, erodes, or bursts after a certain period of the lag phase. In some embodiments, the pharmaceutical composition of this application is formulated for sustained release or delayed sustained release and comprises 100% of the total dose of a particular active agent administered in a unit dosage unit. In other embodiments, the pharmaceutical composition comprises a sustained / delayed release component and an immediate release component. In some embodiments, the immediate release component and the sustained / delayed release component contain the same active ingredient. In other embodiments, the immediate release component and the sustained / delayed release component contain different active ingredients (eg, an analgesic in one component and an antimuscarinic agent in another component). In some embodiments, each of the first and second component contains an analgesic selected from the group consisting of aspirin, ibuprofen, sodium naproxen, indomethacin, nabumetone, and acetaminophen. In other embodiments, the sustained / delayed release component is enteric coated. In other embodiments, the immediate release component and / or the sustained / delayed release component further comprises an antimuscarinic agent selected from the group consisting of oxybutynin, solifenacin, darifenacin and atropine. In other embodiments, the analgesic in each component is administered orally in a daily dose of 5-2000 mg, 20-1000 mg, 50-500 mg, or 250-1000 mg. In other embodiments, the immediate release component and / or the sustained / delayed release component further comprises an antidiuretic agent, an antimuscarinic agent, or both. In other embodiments, the method of treatment comprises administering to the subject a diuretic at least 8 or 7 hours before the target time, such as bedtime, and administering to the subject a pharmaceutical composition comprising an immediate release component and / or a sustained / delayed release component within 2 hours to the target time.

In other embodiments, the “immediate release” component provides about 5-50% of the total dose of the active agent (s) and the “sustained release” component provides 50-95% of the total dose of the active agent (s) delivered by the pharmaceutical. For example, the immediate release component may provide about 20-60%, or about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60% of the total dose of the active agent (s) delivered by the pharmaceutical drug. The sustained release component provides about 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% or 80% of the total dose of the active agent (s) delivered by the pharmaceutical preparation. In some embodiments, the sustained release component further comprises a barrier coating to delay the release of the active agent.

The delayed release barrier coating may consist of many different materials depending on the purpose. In addition, the preparation may contain many barrier coatings to facilitate release at a specific time. The coating may be a sugar coating, a film coating (for example, based on hydroxypropylmethyl cellulose, methyl cellulose, methyl hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, acrylate copolymers, polyethylene glycols and / or polyvinyl pyrrol methyl acetate, hydroxymethyl cellulose acetate succinate, polyvinyl acetate phthalate, shellac and / or ethyl cellulose. In addition, the preparation may further comprise a delay release material, such as, for example, glyceryl monostearate or glyceryl distearate.

In some embodiments, a sustained release formulation comprises an enteric coating comprising one or more polymers that facilitate the release of active agents in the proximal or distal gastrointestinal tract. As used herein, the term “enteric polymer coating” means a coating consisting of one or more polymers having a pH dependent or pH independent release profile. Enteric-coated tablets will not dissolve in acidic gastric juice (pH ~ 3), but will dissolve in an alkaline (pH 7-9) environment of the small intestine or colon. The enteric polymer coating generally prevents the release of active agents before it takes some time after the lag period of gastric emptying, approximately 3-4 hours after administration.

pH-dependent enteric coatings contain one or more pH-dependent or pH-sensitive polymers that retain their structural integrity at low pH, as in the stomach, but dissolve in a higher pH environment in more distal parts of the gastrointestinal tract, such as small intestine, where the contained drug is released. For the purposes of the present invention, “pH dependent” means having characteristics (eg, dissolution) that vary with environmental pH. Exemplary pH-sensitive polymers include, but are not limited to, copolymers of methacrylic acid copolymers, methyl methacrylate-methacrylic acid (e.g., EUDRAGIT ^® L100 (type A), EUDRAGIT ^® S100 (type B), Rohm GmbH, Germany); copolymers of methacrylic acid-ethyl acrylate copolymer (e.g., EUDRAGIT ^® L100-55 (type C) and the dispersion copolymer EUDRAGIT ^® L30D-55, Rohm GmbH, Germany); copolymers of methacrylic acid-methyl methacrylate and methyl methacrylate (EUDRAGIT ^® FS); terpolymers of methacrylic acid, methacrylate and ethyl acrylate; cellulose acetate phthalates (CAP); hydroxypropyl methylcellulose phthalate (HPMCP) (e.g., HP-55, HP-50, HP-55S, Shinetsu Chemical, Japan); polyvinyl acetate phthalate (PVAP) (e.g., COATERIC ^®, OPADRY ^® enteric White OY-P-7171); cellulose acetate succinates (CAS); hydroxypropylmethylcellulose acetate succinate (HPMCAS), e.g., HPMCAS category LF, MF category, HF categories including AQOAT ^® LF and ^{AQOAT ® MF (Shin-Etsu Chemical} , Japan); shellac (e.g. MARCOAT ™ 125 and MARCOAT ™ 125N); carboxymethyl ethyl cellulose (CMEC, Freund Corporation, Japan); cellulose acetate phthalates (CAP) (e.g. AQUATERIC ^® ); cellulose acetate trimellitates (CAT); as well as mixtures of two or more of them in weight ratios of from about 2: 1 to about 5: 1, such as, for example, a mixture of EUDRAGIT ^® L 100-55 and EUDRAGIT ^® S 100 in a weight ratio of about 3: 1 to about 2: 1 or a mixture of EUDRAGIT ^® L 30 D-55 and EUDRAGIT ^® FS in a weight ratio of from about 3: 1 to about 5: 1.

pH-dependent polymers typically have a characteristic optimum pH for dissolution. In some embodiments, the implementation of the pH-dependent polymer has an optimum pH of from about 5.0 to 5.5, from about 5.5 to 6.0, from about 6.0 to 6.5, or from about 6.5 to 7.0 . In other embodiments, the implementation of the pH-dependent polymer has an optimum pH of ≥5.0, ≥5.5, ≥6.0, ≥6.5, or ≥7.0.

These polymers can be used either individually or in combination or together with polymers other than those mentioned above. Preferred enteric pH dependent polymers are pharmaceutically acceptable methacrylic acid copolymers. These copolymers are anionic polymers based on methacrylic acid and methyl methacrylate and preferably have an average molecular weight of about 135,000. The ratio of free carboxyl groups to methyl esterified carboxyl groups in these copolymers can range from, for example, from 1: 1 to 1: 3, for example , approximately 1: 1 or 1: 2. Such polymers are sold under the trade name EUDRAGIT ^® , for example, EUDRAGIT L series, for example, EUDRAGIT L 12.5 ^® , EUDRAGIT L 12.5P ^® , EUDRAGIT L100 ^® , EUDRAGIT L 100-55 ^® , EUDRAGIT L-30D ^® , EUDRAGIT L -30 D-55 ^® , EUDRAGIT S ^® series, e.g. EUDRAGIT S 12.5 ^® , EUDRAGIT S 12.5P ^® , EUDRAGIT S100 ^® . Release promoters are not limited to pH dependent polymers. Other hydrophilic molecules that quickly dissolve and quickly wash out of the dosage form, leaving a porous structure, can also be used for the same purpose.

In a particular embodiment, a coating technique uses mixing one or more pH dependent and one or more pH independent polymers. Mixing pH-dependent and pH-independent polymers can reduce the release rate of the active ingredients after the soluble polymer has reached its optimum pH for dissolution.

In some embodiments, a “time-controlled” or “time-dependent” release profile can be obtained using a water-insoluble capsule containing one or more active agents, the capsule being closed at one end by an insoluble but permeable and swellable hydrogel plug. Upon contact with the gastrointestinal fluid or the dissolution medium, the cork swells, is pushed out of the capsule and the drugs are released after a predetermined lag time period, which can be controlled, for example, by the position and size of the cork. The capsule can be further coated with an external pH-dependent enteric coating, which keeps the capsule intact until it enters the small intestine. Suitable cork materials include, for example, polymethacrylates, erodible compressed polymers (e.g. HPMC, polyvinyl alcohol), solidified molten polymer (e.g. glyceryl monooleate), and enzymatically controlled erodible polymers (e.g. polysaccharides such as amylose, arabinogalactanate, chitonite, , cyclodextrin, dextran, guar gum, pectin and xylan).

In other embodiments, capsules or bilayer tablets may be formulated to include a drug core coated with a swellable layer and an external insoluble but semi-permeable polymer coating or membrane. The lag period to failure can be controlled due to the permeability and mechanical properties of the polymer coating and the nature of the swelling of the swelling layer. Typically, a swellable layer contains one or more swellable substances, such as swellable hydrophilic polymers, which swell and retain water in their structures.

Exemplary, water-swellable materials for use in the coating of delayed release include, but are not limited to, polyethylene oxide (having, for example, an average molecular weight of from 1,000,000 to 7,000,000, such as POLYOX ^®), methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose; polyalkylene oxides having a weight average molecular weight of from 100,000 to 6,000,000, including, but not limited to, poly (methylene oxide), poly (butylene oxide); poly (hydroxyalkyl methacrylate) with a molecular weight of from 25,000 to 5,000,000; poly (vinyl) alcohol with a low acetal content, which is cross-linked with glyoxal, formaldehyde or glutaraldehyde and having a degree of polymerization from 200 to 30,000; mixtures of methyl cellulose, crosslinked agar and carboxymethyl cellulose; hydrogel-forming copolymers obtained by dispersing a finely ground copolymer of maleic anhydride with styrene, ethylene, propylene, butylene or isobutylene, crosslinked with 0.001-0.5 moles of a saturated crosslinking agent per mole of maleic anhydride in the copolymer; acid carboxypolymers CARBOPOL ^® with a molecular weight of 450,000 to 4,000,000; polyacrylamides CYANAMER ^® ; crosslinked water-swellable polymers of indenmaleic anhydride; GOODRITE ^® polyacrylic acid with a molecular weight of 80,000 to 200,000; grafted starch copolymers; AQUA-KEEPS ^® acrylate polymer polysaccharides consisting of condensed glucose units such as crosslinked polyglucan diester; carbomers having a viscosity of 3,000 to 60,000 mPa⋅s in the form of an aqueous solution with a concentration of 0.5% -1% w / v; cellulose ethers, such as hydroxypropyl cellulose, having a viscosity of about 1000-7000 mPa · s in the form of an aqueous solution with a concentration of 1% w / w. (25 ° C); hydroxypropyl methylcellulose having a viscosity of about 1000 or higher, preferably 2500 or higher, up to a maximum of 25000 mPa⋅s in the form of an aqueous solution with a concentration of 2% w / v; polyvinylpyrrolidone having a viscosity of about 300-700 mPa · s in the form of an aqueous solution with a concentration of 10% w / v. at 20 ° C, as well as combinations thereof.

Alternatively, drug release times can be controlled by a lag breakdown period, depending on the equilibrium between the stability and the thickness of the water-insoluble polymer membrane (e.g., ethyl cellulose, EC) containing a given amount of micropores at the bottom of the capsule compartment and a certain amount of swelling excipient, such as low substituted hydroxypropyl cellulose (L-HPC) and sodium glycolate. After oral administration, gastrointestinal fluids penetrate the micropores, causing swelling of the swelling excipients, which creates an internal pressure that separates the capsule components, including the first capsule compartment containing the swellable materials, the second capsule compartment containing the drugs, and the outer cap attached to the first capsule compartment.

The enteric layer may further comprise anti-adhesive agents, such as talc or glyceryl monostearate, and / or plasticizers. The enteric layer may additionally contain one or more plasticizers, including, but not limited to, triethyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, polyethylene glycol, acetylated monoglycerides, glycerin, triacetin, propylene glycol, esters of phthalates, iron nitrate, di-diacetate, , castor oil, sorbitol and dibutylsebacate.

In another embodiment, a delayed release formulation uses a permeable, but insoluble film coating to encapsulate the active ingredient and the osmotic agent. As water from the stomach slowly diffuses through the film to the core, the core swells until the film breaks, while the active ingredients are released. The film coating can be adjusted so that it allows for different water penetration rates or release times.

In another embodiment, a sustained release tablet coating is used in the delayed release formulation so that water penetrates through a controlled opening in the coating until the core disintegrates. When the tablet disintegrates, the contained drug is released immediately or over a longer period of time. These and other methods can be modified to achieve a predetermined lag period before drug release begins.

In another embodiment, the active agents are delivered in a formulation intended to provide both a delayed release and a sustained release (delayed sustained). The term “delayed sustained release” is used herein to refer to a drug formulation that provides a pulsed release of the active agent after a predetermined time or lag period after administration, followed by a sustained release of the active agent.

In some embodiments, the immediate release, sustained release, delayed release, or delayed sustained release preparations comprise an active core consisting of one or more inert particles, each in the form of a ball, granule, pill, granular particle, microcapsule, microsphere, microgranule, nanocapsule or nanosphere with drugs applied to their surface in the form of, for example, a film-forming composition, using, for example, a fluidized bed technique or other methodologies known to those skilled in the art. An inert particle can have a different size, provided that it is large enough to remain poorly soluble. Alternatively, the active core can be obtained by granulation and grinding and / or by extrusion and spheronization of a polymer composition containing a drug.

The amount of drug in the core will depend on the dose that is required, and usually varies from about 5 to 90 mass%. Typically, the polymer coating on the active core will be from about 1 to 50%, based on the weight of the coated particles, depending on the required lag period and type of release profile and / or selected polymers and solvents for the coating. Specialists in this field will be able to choose the appropriate amount of drug for coating or inclusion in the core to achieve the desired dosage. In one embodiment, the inactive core may be a sugar sphere or a buffer crystal or an encapsulated buffer crystal, for example, calcium carbonate, sodium bicarbonate, fumaric acid, tartaric acid, and so on, which changes the microenvironment of the drug to facilitate its release.

In some embodiments, for example, delayed release or delayed sustained release compositions can be prepared by coating particles containing a water-soluble / dispersible drug, such as granules, with a mixture of a water-insoluble polymer and an enteric polymer, while the water-insoluble polymer and the enteric polymer be present in a mass ratio of 4: 1 to 1: 1, and the total weight of the coatings is from 10 to 60 mass%, based on the total weight of the coated granules. Layered drug granules may optionally contain an internal dissolution control ethyl cellulose membrane. The composition of the outer layer, as well as the individual masses of the inner and outer layers of the polymer membrane, are optimized to achieve release profiles with the desired circadian rhythm for a given active substance, which is predicted based on in vitro / in vivo correlations.

In other embodiments, the preparations may contain a mixture of the drug-containing particles of immediate release without controlling the dissolution rate of the polymer membrane and delayed-release granules having, for example, a lag period of 2-4 hours after oral administration, thereby providing a double-pulse release profile.

In some embodiments, the active core is coated with one or more layers controlling the dissolution rate of the polymers to obtain the desired release profiles with or without a lag period. The membrane of the inner layer can significantly control the rate of release of the drug after water or body fluids enter the core, while the membrane of the outer layer can provide the desired lag period (the period without release or with a small release of the drug after water or body fluids enter the core ) The membrane of the inner layer may contain a water-insoluble polymer or a mixture of water-insoluble and water-soluble polymers.

Polymers suitable for the outer membrane, which largely controls the lag period up to 6 hours, may include the enteric polymer described above and a water-insoluble polymer in an amount of from 10 to 50 mass%. The ratio of water-insoluble polymer to enteric polymer may vary from 4: 1 to 1: 2, preferably the polymers are present in a ratio of about 1: 1. The water-insoluble polymer used is typically ethyl cellulose.

Typical water-insoluble polymers include ethyl cellulose, polyvinyl acetate (Kollicoat SR # 0D from BASF), neutral copolymers based on ethyl acrylate and methyl methacrylate, copolymers of acrylic and methacrylic esters with quaternary ammonium groups such as EUDRAGIT ^® NE, RS and RS30D RL or RL30D and the like. Typical water-soluble polymers include low molecular weight HPMC, HPC, methyl cellulose, polyethylene glycol (PEG with a molecular weight> 3000) with a layer thickness ranging from 1 mass% up to 10 mass%, depending on the solubility of the active substance in water and the solvent-based coating preparation used or latex suspension. The ratio of water-insoluble polymer to water-soluble polymer may typically vary from 95: 5 to 60:40, preferably from 80:20 to 65:35.

In some embodiments, an AMBERLITE ™ IRP69 resin is used as a sustained release carrier. AMBERLITE ™ IRP69 is an insoluble, strongly acidic sodium form of cation exchange resin, which is suitable as a carrier for cationic (basic) substances. In other embodiments, a DUOLITE ™ AP143 / 1093 resin is used as a sustained release carrier. DUOLITE ™ AP143 / 1093 is an insoluble, highly alkaline anion-exchange resin, which is suitable as a carrier for anionic (acidic) substances.

When used as a drug carrier, AMBERLITE IRP69 or / and DUOLITE ™ AP143 / 1093 resin provides the ability to bind drugs on an insoluble polymer matrix. Sustained release is achieved through the formation of resin and drug complexes (drug resinates). The drug is released from the resin in vivo when the drug reaches equilibrium with high concentrations of electrolytes, which are typical of the gastrointestinal tract. More hydrophobic drugs will typically elute from the resin at a lower rate due to hydrophobic interaction with the aromatic structure of the cation exchange system.

In some embodiments, the pharmaceutical composition is formulated for oral administration. Oral dosage forms include, for example, tablets, capsules, caplets, and may also contain many granules, beads, powders, or pills, which may or may not be encapsulated. Tablets and capsules represent the most convenient oral dosage forms, in which case solid pharmaceutical carriers are used.

In a delayed release preparation, one or more barrier coatings may be applied to pills, tablets, or capsules to facilitate slow dissolution and concomitant release of drugs into the intestines. Typically, the barrier coating contains one or more polymers enclosing, surrounding or forming a layer or membrane around a therapeutic composition or active nucleus.

In some embodiments, the active agents are delivered in a formulation intended to provide a delayed release at a predetermined time after administration. This delay can be up to about 10 minutes, about 20 minutes, about 30 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours or longer.

Various coating methods can be applied to granules, beads, powders or pills, tablets, capsules, or a combination thereof containing active agents, to obtain different and different release profiles. In some embodiments, the pharmaceutical composition is in the form of a tablet or capsule containing one coating layer. In other embodiments, the pharmaceutical composition is in the form of a tablet or capsule containing several coating layers.

In some embodiments, the pharmaceutical composition comprises several active ingredients selected from the group consisting of analgesics, antimuscarinic agents, antidiuretics, antispasmodics, and zolpidem. Examples of antimuscarinic agents include, but are not limited to, oxybutynin, solifenacin, darifenacin, and atropine. Examples of antidiuretics include, but are not limited to, antidiuretic hormone (ADH), angiotensin II, aldosterone, vasopressin, vasopressin analogs (eg, desmopressin, argipressin, lipressin, felipressin, ornipressin, terlipressin); vasopressin receptor agonists, atrial natriuretic peptide (ANP) receptor antagonists, and C-type natriuretic peptide (CNP) (i.e., NPR1, NPR2, NPR3) (e.g., HS-142-1, isatin [Asu7.23 '] b-ANP- (7-28)], anantine, a cyclic peptide from Streptomyces coerulescens, and monoclonal antibody 3G12); type 2 somatostatin receptor antagonists (e.g., somatostatin), as well as their pharmaceutically acceptable derivatives, analogs, salts, hydrates and solvates. Examples of antispasmodics include, but are not limited to, carisoprodol, benzodiazepines, baclofen, cyclobenzaprine, metaxalone, metocarbamol, clonidine, an analogue of clonidine and dantrolene. In some embodiments, the pharmaceutical composition comprises one or more analgesics. In other embodiments, the pharmaceutical composition comprises (1) one or more analgesics and (2) one or more other active ingredients selected from the group consisting of antimuscarinic agents, antidiuretics, and antispasmodics. In another embodiment, the pharmaceutical composition comprises (1) one or more analgesics and (2) one or more antimuscarinic agents. In another embodiment, the pharmaceutical composition comprises (1) one or more analgesics and (2) one or more antidiuretics. In another embodiment, the pharmaceutical composition comprises (1) one or more analgesics and (2) one or more antispasmodics. In another embodiment, the pharmaceutical composition comprises (1) one or more analgesics and (2) zolpidem. In another embodiment, the pharmaceutical composition comprises (1) one or two analgesics, (2) one or two antimuscarinic agents, and (3) one or two antidiuretics. In another embodiment, the pharmaceutical composition comprises (1) one or more analgesics, (2) one or more antispasmodics, and (3) one or more antidiuretics. In another embodiment, the pharmaceutical composition comprises (1) one or more analgesics, (2) one or more antidiuretics, and (3) zolpidem.

In one embodiment, several active ingredients are formulated for immediate release. In another embodiment, several active ingredients are formulated for sustained release. In another embodiment, several active ingredients are formulated for both immediate release and sustained release (for example, a first portion of each active ingredient is formulated for immediate release and a second portion of each active ingredient is formulated for sustained release). In another embodiment, some of the several active ingredients are formulated for immediate release and some of the several active ingredients are formulated for sustained release (for example, active ingredients A, B, C are formulated for immediate release, and active ingredients C and D are formulated for sustained release). In some other embodiments, the immediate release component and / or the sustained release component is further coated with a delayed release coating such as an enteric coating.

In some embodiments, the pharmaceutical composition comprises an immediate release component and a sustained release component. The immediate release component may contain one or more active ingredients selected from the group consisting of analgesics, antimuscarinic agents, antidiuretics, and antispasmodics. The sustained release component may contain one or more active ingredients selected from the group consisting of analgesics, antimuscarinic agents, antidiuretics, and antispasmodics. In some embodiments, the immediate release component and the sustained release component have exactly the same active ingredients. In other embodiments, the immediate release component and the sustained release component have different active ingredients. In other embodiments, the immediate release component and the sustained release component have one or more common active ingredients. In some other embodiments, the immediate release component and / or the sustained release component is further coated with a delayed release coating such as an enteric coating.

In one embodiment, the pharmaceutical composition comprises two or more active ingredients (for example, two or more analgesic agents or a mixture of one or more analgesic agents and one or more antimuscarinic agents or antidiuretics or antispasmodics or zolpidem) formulated for immediate release in about one and same time. In another embodiment, the pharmaceutical composition comprises two or more active ingredients formulated for sustained release at about the same time. In another embodiment, the pharmaceutical composition comprises two or more active ingredients formulated as two sustained release components, each with a different sustained release profile. For example, the first sustained release component releases the first active ingredient at a first release rate and the second sustained release component releases a second active ingredient at a second release rate. In another embodiment, the pharmaceutical composition comprises two or more active ingredients, both formulated for delayed release. In another embodiment, the pharmaceutical composition comprises two or more active ingredients formulated for delayed release. In another embodiment, the pharmaceutical composition comprises two or more active ingredients formulated as two delayed release components, each with a different delayed release profile. For example, the first delayed release component releases the first active ingredient at the first point in time and the second delayed release component releases the second active ingredient at the second point in time. In another embodiment, the pharmaceutical composition comprises two or more active ingredients, one or more of which are formulated for immediate release, and the other are formulated for sustained release. In another embodiment, the pharmaceutical composition comprises two or more active ingredients, some of which are formulated for immediate release and the remainder are formulated for sustained release.

In other embodiments, the pharmaceutical composition comprises two active ingredients (e.g., two analgesic agents, or a mixture of one analgesic and one antimuscarinic agent or antidiuretic, or antispasmodic, or zolpidem) formulated for immediate release, and (2) two active ingredients ( for example, two analgesic agents, or a mixture of one analgesic and one antimuscarinic or antidiuretic, or antispasmodic, or zolpidem), compiled for prolonged release. In other embodiments, the pharmaceutical composition comprises three active ingredients formulated for immediate release, and (2) three active ingredients formulated for sustained release. In other embodiments, the pharmaceutical composition comprises four active ingredients formulated for immediate release and (2) four active ingredients formulated for sustained release. In these embodiments, the active ingredient (s) in the immediate release component may be the same or different from the active ingredient (s) in the sustained release component. In some other embodiments, the immediate release component and / or the sustained release component is further coated with a delayed release coating such as an enteric coating.

In some embodiments, the pharmaceutical composition comprises one or more analgesic agents and an antidiuretic, wherein one or more analgesic agents are formulated for delayed release and the antidiuretic is formulated for immediate release. In other embodiments, the pharmaceutical composition further comprises an additional agent selected from the group consisting of an antimuscarinic agent, antidiuretic agent, antispasmodic, and zolpidem, wherein the additional agent is formulated for delayed release. In some embodiments, the delayed release formulation delays the release of the active ingredient (eg, analgesic, antimuscarinic, antidiuretic, antispasmodic and / or zolpidem) for 1, 2, 3, 4 or 5 hours.

The term “immediate release” is used herein to refer to a drug formulation that does not contain dissolution rate controlling material. There is practically no delay in the release of active agents after administration of the immediate release preparation. The immediate release coating may contain suitable materials that dissolve immediately after administration to release the drug contained therein. Exemplary coating materials for immediate release include gelatin, polyvinyl alcohol copolymers and polyethylene glycol (PVA-PEG) (e.g., KOLLICOAT ^® and various other materials known to those skilled in the art.

The immediate release composition may contain 100% of the total dose of a particular active agent administered in a unit dosage unit. Alternatively, the immediate release component can be included as a component in a preparation with a combined release profile, which can ensure that the pharmaceutical preparation delivers from about 1% to about 60% of the total dose of the active agent (s). For example, an immediate release component may provide drug delivery of about 5% -60%, from about 10% to about 60%, from about 10% to about 50%, from about 10% to about 40%, from about 10% to about 30 %, from about 10% to about 20%, from about 20% to about 60%, from about 20% to about 50%, from about 20% to about 30%, from about 30% to about 60%, from about 30 % to about 50%, from about 40% to about 60%, from about 40% to about 50%, from about 45% to about 60%, or from about 45% to about 50% of the total dose of the active agent (medium stem). In alternative embodiments, the immediate release component provides a drug delivery of about 2, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, or 60% of the total dose of the active agent (s).

In some embodiments, the immediate release or delayed release preparation comprises an active core consisting of one or more inert particles, each in the form of a ball, granule, pill, granular particle, microcapsule, microsphere, microgranule, nanocapsule or nanosphere, coated with drug means in the form of, for example, a drug-containing film-forming composition, using, for example, a fluidized bed technique or other methodologies known Professionals widely in this area. An inert particle can have a different size, provided that it is large enough to remain poorly soluble. Alternatively, the active core can be obtained by granulation and grinding and / or by extrusion and spheronization of a polymer composition containing a drug.

The amount of drug in the core will depend on the dose that is required, and usually varies from about 5 to 90 mass%. Typically, the polymer coating on the active core will be from about 1 to 50%, based on the weight of the coated particles, depending on the required lag period and type of release profile and / or selected polymers and solvents for the coating. Specialists in this field will be able to choose the appropriate amount of drug for coating or inclusion in the core to achieve the desired dosage. In one embodiment, the inactive core may be a sugar sphere or buffer crystal, or an encapsulated buffer crystal, for example, calcium carbonate, sodium bicarbonate, fumaric acid, tartaric acid, and so on, which changes the microenvironment of the drug to facilitate its release.

In some embodiments, a delayed release preparation is prepared by coating particles containing a water-soluble / dispersible drug, such as granules, with a mixture of a water-insoluble polymer and an enteric polymer, while the water-insoluble polymer and the enteric polymer may be present in a weight ratio of 4: 1 up to 1: 1 and the total weight of the coatings is from 10 to 60 mass%, calculated on the total weight of the coated granules. Layered drug granules may optionally contain an internal dissolution control ethyl cellulose membrane. The composition of the outer layer, as well as the individual masses of the inner and outer layers of the polymer membrane, are optimized to achieve release profiles with the desired circadian rhythm for a given active substance, which is predicted based on in vitro / in vivo correlations.

In other embodiments, the preparations comprise a mixture of drug-containing particles of immediate release without controlling the dissolution rate of the polymer membrane and delayed-release granules having, for example, a lag period of 2-4 hours after oral administration, thereby providing a dual-pulse release profile. In other embodiments, the preparations comprise a mixture of two types of delayed release granules: the first type having a lag period of 1-3 hours, and the second type having a lag period of 4-6 hours.

Preferably, the formulations are designed with release profiles such as to limit obstacles to proper sleep, and the drug is released from the formulation at a time when the person would normally wake up from urge to urinate. For example, consider the case when a person falls asleep at 11 a.m. and usually wakes up at 12:30 a.m., 3:00 a.m. and 6:00 a.m. to urinate. A delayed sustained release vehicle may be taken at 10 p.m. and will begin to deliver the drug at 12 a.m. and gradually release the drug within 5-8 hours, thereby delaying or eliminating the need to urinate.

In other embodiments, the preparations are formulated with release profiles such that a portion of the drug (e.g., 20-60%) is released immediately or within 2 hours after administration and the residue is released over an extended period of time. The pharmaceutical composition can be administered daily or administered as needed. In some embodiments, the pharmaceutical composition is administered to a subject before bedtime. In some embodiments, the pharmaceutical composition is administered immediately before bedtime. In some embodiments, the pharmaceutical composition is administered within about two hours before bedtime, preferably within about one hour before bedtime. In another embodiment, the pharmaceutical composition is administered about two hours before bedtime. In a further embodiment, the pharmaceutical composition is administered at least two hours before bedtime. In yet another embodiment, the pharmaceutical composition is administered about one hour before bedtime. In a further embodiment, the pharmaceutical composition is administered at least one hour before bedtime. In yet another embodiment, the pharmaceutical composition is administered less than one hour before bedtime. In another embodiment, the pharmaceutical composition is administered immediately before bedtime. Preferably, the pharmaceutical composition is administered orally.

The appropriate dose ("therapeutically effective amount") of the active agent (s) in the immediate release component or the sustained release component will depend, for example, on the severity and course of the condition, route of administration, bioavailability of the specific agent (s), age and body weight of the patient, history the patient’s illness and reactions to the active agent (s), the opinions of the doctor and so on.

As a general suggestion, a therapeutically effective amount of the active agent (s) to be administered in the immediate release component, sustained release component or delayed sustained release component is in the range of about 100 μg / kg body weight / day to about 100 mg / kg body weight / day either for one or for several introductions. In some embodiments, the range for each active agent administered daily in a single dose or in multiple doses is from about 100 μg / kg body weight / day to about 50 mg / kg body weight / day, from 100 μg / kg body weight / day to about 10 mg / kg body weight / day, from 100 μg / kg body weight / day to about 1 mg / kg body weight / day, from 100 μg / kg body weight / day to about 10 mg / kg body weight / day, from 500 μg / kg body weight / day to about 100 mg / kg body weight / day, from 500 μg / kg body weight / day to about 50 mg / kg body weight / day, from 500 μg / kg body weight / day to about 5 mg / kg body weight / day, from 1 mg / kg body weight / day to about 100 mg / kg body weight / day, from 1 mg / kg body weight / day to about 50 mg / kg body weight / day, from 1 mg / kg body weight / day to about 10 mg / kg body weight / day, from 5 mg / kg body weight / day to about 100 mg / kg body weight / day, from 5 mg / kg body weight / day to about 50 mg / kg body weight / day, from 10 mg / kg body weight / day to about 100 mg / kg body weight / day and from 10 mg / kg body weight / day to approximately 50 mg / kg body weight / day.

The active agent (a) described herein can be included in an immediate release component or a sustained release component, a delayed sustained release component, or a combination thereof for daily oral administration in a single dose or combined dose in the range of 1 mg to 2000 mg, 5 mg up to 2000 mg, from 10 mg to 2000 mg, from 50 mg to 2000 mg, from 100 mg to 2000 mg, from 200 mg to 2000 mg, from 500 mg to 2000 mg, from 5 mg to 1800 mg, from 10 mg to 1600 mg, 50 mg to 1600 mg, 100 mg to 1500 mg, 150 mg to 1200 mg, 200 mg to 1000 mg, 300 mg to 800 mg, 325 mg to 500 mg, 1 mg to 1000 mg, 1 mg to 500 mg, 1 mg to 200 mg, 5 mg to 1000 mg, 5 mg to 500 mg, 5 mg to 200 mg, 10 mg to 1000 mg , from 10 mg to 500 mg, from 10 mg to 200 mg, from 50 mg to 1000 mg, from 50 mg to 500 mg, from 50 mg to 200 mg, from 250 mg to 1000 mg, from 250 mg to 500 mg, 500 mg to 1000 mg, 500 mg to 2000 mg. As expected, the dose will depend on the pathological condition, size, age and health status of the patient.

In some embodiments, the pharmaceutical composition comprises one analgesic. In one embodiment, one analgesic agent is aspirin. In another embodiment, one analgesic agent is ibuprofen. In another embodiment, one analgesic is naproxen or naproxen sodium. In another embodiment, one analgesic agent is indomethacin. In another embodiment, one analgesic agent is nabumetone. In another embodiment, one analgesic agent is acetaminophen.

In some embodiments, a single analgesic is administered at a daily dose of 1 mg to 2000 mg, 5 mg to 2000 mg, 20 mg to 2000 mg, 5 mg to 1000 mg, 20 mg to 1000 mg, 50 mg to 500 mg, 100 mg to 500 mg, 250 mg to 500 mg, 250 mg to 1000 mg, or 500 mg to 1000 mg. In some embodiments, the pharmaceutical composition comprises acetylsalicylic acid, ibuprofen, naproxen, naproxen sodium, indomethacin, nabumetone or acetaminophen as a single analgesic agent and the analgesic agent is administered orally in a daily dose ranging from 5 mg to 2000 mg, from 20 mg to 2000 mg, from 5 mg to 1000 mg, from 20 mg to 1000 mg, from 50 mg to 500 mg, from 100 mg to 500 mg, from 250 mg to 500 mg, from 250 mg to 1000 mg or from 500 mg to 1000 mg In some embodiments, the second analgesic is administered in a daily dose of 1 mg to 2000 mg, 5 mg to 2000 mg, 20 mg to 2000 mg, 5 mg to 1000 mg, 20 mg to 1000 mg, 50 mg to 500 mg, 100 mg to 500 mg, 250 mg to 500 mg, 250 mg to 1000 mg, or 500 mg to 1000 mg.

In other embodiments, the pharmaceutical composition comprises a pair of analgesic agents. Examples of such pairs of analgesics include but are not limited to, acetylsalicylic acid, ibuprofen, acetylsalicylic acid and sodium naproxen, acetylsalicylic acid, nabumetone, aspirin and acetaminophen, acetylsalicylic acid, indomethacin, ibuprofen and naproxen sodium, ibuprofen, nabumetone, ibuprofen and acetaminophen , ibuprofen and indomethacin, naproxen, naproxen sodium and nabumetone, naproxen sodium and acetaminophen, naproxen sodium and indomethacin, nabumetone and acetaminophen, nabumetone and indome tacin, as well as acetaminophen and indomethacin. Pairs of analgesic agents are mixed in a mass ratio in the range from 0.1: 1 to 10: 1, from 0.2: 1 to 5: 1, or from 0.3: 1 to 3: 1, while the combined dose is in range from 5 mg to 2000 mg, from 20 mg to 2000 mg, from 100 mg to 2000 mg, from 200 mg to 2000 mg, from 500 mg to 2000 mg, from 5 mg to 1500 mg, from 20 mg to 1500 mg, from 100 mg to 1500 mg, from 200 mg to 1500 mg, from 500 mg to 1500 mg, from 5 mg to 1000 mg, from 20 mg to 1000 mg, from 100 mg to 1000 mg, from 250 mg to 500 mg, 250 mg to 1000 mg, from 250 mg to 1500 mg, from 500 mg to 1000 mg, from 500 mg to 1500 mg, from 1000 mg to 1500 mg and from 1000 mg to 2000 mg. In one embodiment, pairs of analgesic agents are mixed in a weight ratio of 1: 1.

In some other embodiments, the pharmaceutical composition of this application further comprises one or more antimuscarinic agents. Examples of antimuscarinic agents include, but are not limited to, oxybutynin, solifenacin, darifenacin, fesoterodine, tolterodine, trospium, and atropine. The daily dose of an antimuscarinic agent is in the range from 0.01 mg to 100 mg, from 0.1 mg to 100 mg, from 1 mg to 100 mg, from 10 mg to 100 mg, from 0.01 mg to 25 mg, from 0 , 1 mg to 25 mg, from 1 mg to 25 mg, from 10 mg to 25 mg, from 0.01 mg to 10 mg, from 0.1 mg to 10 mg, from 1 mg to 10 mg, from 10 mg to 100 mg and 10 mg to 25 mg.

In some embodiments, the pharmaceutical composition comprises an analgesic agent selected from the group consisting of acetylsalicylic acid, ibuprofen, naproxen, naproxen sodium, nabumetone, acetaminophen and indomethacin, and an antimuscarinic agent selected from the group consisting of oxybutynin, solifenacin, and dari.

Another aspect of the present application relates to a method for decreasing the frequency of urination by administering to a person in need thereof a pharmaceutical composition formulated in an immediate release preparation. The pharmaceutical composition contains several analgesic agents and / or antimuscarinic agents.

In some embodiments, the pharmaceutical composition comprises two or more analgesic agents. In other embodiments, the pharmaceutical composition comprises one or more analgesic agents and one or more antimuscarinic agents. The pharmaceutical composition may be in the form of a tablet, capsule, dragee, powder, granulate, liquid, gel or emulsion. The specified liquid, gel or emulsion, the subject can take in pure form or contained inside the capsule.

In some embodiments, the analgesic is selected from the group consisting of salicylates, aspirin, salicylic acid, methyl salicylate, diflunisal, salsalate, olsalazine, sulfasalazine, derivatives of paraaminophenol, acetanilide, acetaminophen, phenacetin, phenamates, mefenamefenamic acid, mefenamefenamic acid acetic acid, tolmetin, ketorolac, diclofenac, derivatives of propionic acid, ibuprofen, sodium naproxen, naproxen, phenoprofen, ketoprofen, flurbiprof hen, oxaprozin; enolic acids, derivatives of oxicam, piroxicam, meloxicam, tenoxicam, ampiroxicam, droxicam, pivoxicam, derivatives of pyrazolone, phenylbutazone, oxyphenbutazone, antipyrine, aminopyrine, dipyrone, coxibam, celecoxib, rofecoxin apolumide, indufinomene indubene, nabumetin, indubene, nabuodimene, nabu-indomene, nabu-indomene, nabu-indomene, nabu-indomene, and isobutylphenylpropionic acid. The antimuscarinic agent is selected from the group consisting of oxybutynin, solifenacin, darifenacin and atropine.

In some embodiments, the pharmaceutical composition comprises one analgesic and one antimuscarinic. In one embodiment, one analgesic agent is aspirin. In another embodiment, one analgesic agent is ibuprofen. In another embodiment, one analgesic is naproxen or naproxen sodium. In another embodiment, one analgesic agent is indomethacin. In another embodiment, one analgesic agent is nabumetone. In another embodiment, one analgesic agent is acetaminophen. The analgesic and antimuscarinic can be taken in doses in the ranges indicated above.

In some embodiments, the pharmaceutical composition comprises one or more analgesic agents, individually or in combination, in an amount in the range of 50-2000 mg, 50-1500 mg, 50-1200 mg, 50-1000 mg, 50-800 mg, 50-600 mg, 50-500 mg, 50-400 mg, 50-300 mg, 50-250 mg, 50-200 mg, 50-150 mg, 50-100 mg, 100-2000 mg, 100-1500 mg, 100-1200 mg, 100-1000 mg, 100-800 mg, 100-600 mg, 100-400 mg, 100-250 mg, 250-2000 mg, 250-1500 mg, 250-1200 mg, 250-1000 mg, 250-800 mg, 250-600 mg, 250-400 mg, 400-2000 mg, 400-1500 mg, 400-1200 mg, 400-1000 mg, 400-800 mg, 400-600 mg, 600-2000 mg, 600-1500 mg, 600-1200 mg, 600-1000 mg, 600-800 mg, 800-2000 mg, 800-1500 mg, 800-1200 mg, 800-1000 mg, 1000-2000 mg, 1000-1500 mg, 1000-1200 mg, 1200-2000 mg, 1200-15 00 mg or 1500-2000 mg, wherein the composition is formulated for sustained release with a release profile in which one or more analgesic agents are released continuously for 5-24 hours, 5-8, 8-16 hours or 16-24 hours.

In some embodiments, the composition is formulated for sustained release with a release profile in which at least 90% of one or more analgesic agents are released continuously for 5-24 hours, 5-8, 8-16 hours, or 16-24 hours.

In some embodiments, the composition is formulated for sustained release with a release profile in which one or more analgesic agents are released continuously for 5, 6, 7, 8, 10, 12, 14, 16, 18, 20, 22, or 24 hours. In some embodiments, the pharmaceutical composition further comprises an antimuscarinic agent, antidiuretic agent, or antispasmodic.

In other embodiments, the composition is formulated for sustained release with a release profile in which the analgesic is released at a constant rate over 5-24 hours, 5-8, 8-16 hours, or 16-24 hours. In other embodiments, the composition is formulated for sustained release with a release profile in which the analgesic is released at a constant rate over 5, 6, 7, 8, 10, 12, 14, 16, 18, 20, 22, or 24 hours. As used herein, the expression “at a constant rate over a given period of time” means a release profile in which the release rate at any time over a given period of time is within 30-300% of the average release rate during this predetermined period of time. For example, if 80 mg of aspirin is released at a constant rate over 8 hours, the average release rate is 10 mg / hour during this time period and the actual release rate at any time during this period is in the range of 3 mg / hour to 30 mg / hour (i.e., within 30-300% of the average release rate of 10 mg / hour over an 8-hour period). In some embodiments, the pharmaceutical composition further comprises an antimuscarinic agent, antidiuretic agent, or antispasmodic.

In some embodiments, the analgesic is selected from the group consisting of aspirin, ibuprofen, sodium naproxen, naproxen, indomethacin, nabumetone, and acetaminophen. The pharmaceutical composition is formulated to provide stable release of a small amount of an analgesic to maintain an effective concentration of the drug in the blood, and therefore the total amount of the drug in one dose is reduced compared to the immediate release preparation.

In some embodiments, the pharmaceutical composition comprises 50-250 mg, 250-400 mg, or 400-600 mg of an analgesic formulated for sustained release, with a release profile in which at least 90% of the analgesic is released continuously or at a constant rate over a period of 5-24, 5-8, 8-16 or 16-24 hours.

In one particular embodiment, the pharmaceutical composition comprises 50-250 mg of acetaminophen formulated for sustained release, with a release profile in which at least 90% of acetaminophen is released continuously or at a constant rate over 5-24, 5-8, 8-16 or 16-24 hours.

In another specific embodiment, the pharmaceutical composition comprises 250-400 mg of acetaminophen formulated for sustained release, with a release profile in which 90% of acetaminophen is released continuously or at a constant rate for 5-24, 5-8, 8-16, or 16- 24 hours.

In another specific embodiment, the pharmaceutical composition comprises 400-600 mg of acetaminophen formulated for sustained release, with a release profile in which 90% of acetaminophen is released continuously or at a constant rate for 5-24, 5-8, 8-16, or 16- 24 hours.

In another specific embodiment, the pharmaceutical composition comprises 600-800 mg of acetaminophen formulated for sustained release, with a release profile in which 90% of acetaminophen is released continuously or at a constant rate for 5-24, 5-8, 8-16, or 16- 24 hours.

In another embodiment, the pharmaceutical composition comprises 800-1000 mg of acetaminophen formulated for sustained release, with a release profile in which 90% of acetaminophen is released continuously or at a constant rate for 5-24, 5-8, 8-16, or 16-24 hours.

In some other embodiments, the pharmaceutical composition comprises one or more analgesic agents, individually or in combination, in an amount in the range of 50-2000 mg, 50-1500 mg, 50-1200 mg, 50-1000 mg, 50-800 mg, 50- 600 mg, 50-500 mg, 50-400 mg, 50-300 mg, 50-250 mg, 50-200 mg, 100-2000 mg, 100-1500 mg, 100-1200 mg, 100-1000 mg, 100- 800 mg, 100-600 mg, 100-500 mg, 100-400 mg, 100-300 mg, 100-200 mg, 200-2000 mg, 200-1500 mg, 200-1200 mg, 200-1000 mg, 200- 800 mg, 200-600 mg, 200-400 mg, 400-2000 mg, 400-1500 mg, 400-1200 mg, 400-1000 mg, 400-800 mg, 400-600 mg, 600-2000 mg, 600- 1500 mg, 600-1200 mg, 600-1000 mg, 600-800 mg, 800-2000 mg, 800-1500 mg, 800-1200 mg, 800-1000 mg, 1000-2000 mg, 1000-1500 mg, 1000- 1200 mg, 1200-200 0 mg, 1200-1500 mg or 1500-2000 mg, wherein the analgesic agent (a) is formulated for sustained release, characterized by a two-phase release profile in which 20-60% of the analgesic agent (s) is released within 2 hours after administration and the remainder released continuously or at a constant rate for 5-24 hours. In another embodiment, the analgesic agent (a) is formulated for sustained release with a biphasic release profile in which 20, 30, 40, 50, or 60% of the analgesic agent (s) is released within 2 hours after administration and the residue is released continuously or at a constant rate within 5-8, 8-16 or 16-24 hours. In one embodiment, the analgesic agent (a) is selected from the group consisting of aspirin, ibuprofen, sodium naproxen, naproxen, indomethacin, nabumetone and acetaminophen. In another embodiment, the analgesic is acetaminophen. In some embodiments, the pharmaceutical composition further comprises an antimuscarinic agent, antidiuretic agent, antispasmodic, or zolpidem.

In another embodiment, the pharmaceutical composition comprises 50-400 mg of acetaminophen formulated for sustained release, with a biphasic release profile in which 20%, 30%, 40%, 50%, or 60% of acetaminophen is released within 2 hours after administration and the residue is released continuously or at a constant speed for 5-24, 5-8, 8-16 or 16-24 hours.

In another embodiment, the pharmaceutical composition comprises 100-300 mg of acetaminophen formulated for sustained release, with a biphasic release profile in which 20%, 30%, 40%, 50%, or 60% of acetaminophen is released within 2 hours after administration and the residue is released continuously or at a constant speed for 5-24, 5-8, 8-16 or 16-24 hours.

In another embodiment, the pharmaceutical composition comprises 400-600 mg of acetaminophen formulated for sustained release with a biphasic release profile in which 20%, 30%, 40%, 50%, or 60% of acetaminophen is released within 2 hours after administration and the residue is released continuously or at a constant speed for 5-24, 5-8, 8-16 or 16-24 hours.

In another embodiment, the pharmaceutical composition contains 600-800 mg of acetaminophen formulated for sustained release, with a biphasic release profile in which 20%, 30%, 40%, 50%, or 60% of acetaminophen is released within 2 hours after administration and the residue is released continuously or at a constant speed for 5-24, 5-8, 8-16 or 16-24 hours.

In another embodiment, the pharmaceutical composition comprises 800-1000 mg of acetaminophen formulated for sustained release, with a biphasic release profile in which 20%, 30%, 40%, 50%, or 60% of acetaminophen is released within 2 hours after administration and the residue is released continuously or at a constant speed for 5-24, 5-8, 8-16 or 16-24 hours.

In another embodiment, the pharmaceutical composition comprises 1000-1200 mg of acetaminophen formulated for sustained release with a biphasic release profile in which 20%, 30%, 40%, 50%, or 60% of acetaminophen is released within 2 hours after administration and the residue is released continuously or at a constant speed for 5-24, 5-8, 8-16 or 16-24 hours.

Another aspect of the present application relates to a method for treating nocturia by administering to a person in need thereof a first pharmaceutical composition comprising a diuretic, followed by administration of a second pharmaceutical composition containing one or more analgesic agents. The first pharmaceutical composition is dosed and formulated to provide a diuretic effect within 6 hours after administration and is administered at least 8 or 7 hours before bedtime. A second pharmaceutical composition is formulated for sustained release or delayed sustained release and is administered within 2 hours of sleep.

Examples of diuretics include, but are not limited to, acidifying salts such as CaCl ₂ and NH ₄ Cl; arginine-vasopressin receptor 2 antagonists such as amphotericin B and lithium citrate; watercolors such as Goldenrod and Junipe; Na-H exchange antagonists such as dopamine; carbonic anhydrase inhibitors such as acetazolamide and dorzolamide; loop diuretics such as bumetanide, ethacrylic acid, furosemide and torsemide; osmotic diuretics such as glucose and mannitol; potassium-preserving diuretics such as amiloride, spironolactone, triamteren, potassium canrenoate; thiazides, such as bendroflumethiazide and hydrochlorothiazide, and xanthines, such as caffeine, theophylline and theobromine.

In some embodiments, the second pharmaceutical composition further comprises one or more antimuscarinic agents. In some other embodiments, the second pharmaceutical composition further comprises one or more antidiuretic agents. In some other embodiments, the second pharmaceutical composition further comprises one or more antispasmodics. In some other embodiments, the second pharmaceutical composition further comprises zolpidem. The second pharmaceutical composition may be formulated into an immediate release preparation or a delayed release preparation.

Another aspect of the present application relates to a method of reducing the frequency of urination by administering to a subject in need of two or more analgesic agents alternately to prevent the development of drug resistance. In one embodiment, the method comprises administering a first analgesic agent in a first time period and then administering a second analgesic agent in a second time period. In another embodiment, the method further comprises administering a third analgesic agent in a third time period. The first, second and third analgesic agents are different from each other and at least one of them is formulated for sustained release or delayed sustained release. In one embodiment, the first analgesic is acetaminophen, the second analgesic is ibuprofen, and the third analgesic is naproxen sodium. The duration of each period may vary depending on the subject's response to each analgesic. In some embodiments, each period lasts from 3 days to three weeks. In another embodiment, the first, second, and third analgesics are formulated for sustained release or delayed sustained release.

Another aspect of the present application relates to a pharmaceutical composition comprising several active ingredients and a pharmaceutically acceptable carrier, wherein at least one of the several active ingredients is formulated for sustained release or delayed sustained release. In some embodiments, several active ingredients include one or more analgesics and one or more antidiuretics. In other embodiments, several active ingredients include one or more analgesics and one or more antimuscarinic agents. In other embodiments, several active ingredients include one or more analgesics and zolpidem. In other embodiments, several active ingredients include one or more analgesics, one or more antidiuretic agents, and one or more antimuscarinic agents. In other embodiments, several active ingredients include one or more analgesics, zolpidem, and one or more antidiuretic agents or one or more antimuscarinic agents. The antimuscarinic agent may be selected from the group consisting of oxybutynin, solifenacin, darifenacin and atropine. In other embodiments, the pharmaceutical composition comprises two different analgesics selected from the group consisting of acetylsalicylic acid, ibuprofen, sodium naproxen, naproxen, nabumetone, acetaminophen and indomethacin. In other embodiments, the pharmaceutical composition comprises one analgesic selected from the group consisting of acetylsalicylic acid, ibuprofen, naproxen sodium, nabumetone, acetaminophen, and indomethacin; and an antimuscarinic agent selected from the group consisting of oxybutynin, solifenacin, darifenacin and atropine.

In other embodiments, the pharmaceutical composition of the present application further comprises one or more antispasmodics. Examples of antispasmodics include, but are not limited to, carisoprodol, benzodiazepines, baclofen, cyclobenzaprine, metaxalone, metocarbamol, clonidine, an analogue of clonidine and dantrolene. In some embodiments, antispasmodics are used in a daily dose of from 1 mg to 1000 mg, from 1 mg to 100 mg, from 10 mg to 1000 mg, from 10 mg to 100 mg, from 20 mg to 1000 mg, from 20 mg to 800 mg , from 20 mg to 500 mg, from 20 mg to 200 mg, from 50 mg to 1000 mg, from 50 mg to 800 mg, from 50 mg to 200 mg, from 100 mg to 800 mg, from 100 mg to 500 mg, 200 mg to 800 mg and 200 mg to 500 mg. Antispasmodics may be formulated alone or together with other active ingredient (s) in a pharmaceutical composition for immediate release, sustained release, delayed sustained release, or a combination thereof.

In some embodiments, the pharmaceutical composition comprises one or more analgesic agents selected from the group consisting of aspirin, ibuprofen, naproxen, naproxen sodium, indomethacin, nabumetone and acetaminophen in an amount of 50-400 mg for each agent, and one or more antimuscarinic agents, selected from the group consisting of oxybutynin, solifenacin, darifenacin and atropine in a total amount of 1-25 mg, the pharmaceutical composition being formulated for sustained release with a biphasic profile in release, in which 20-60% of the active ingredients are released within 2 hours after administration and the remainder of the active ingredients is released continuously or at a constant rate for 5-24 hours, 5-8 hours, 8-16 hours or 16-24 hours.

In some embodiments, the pharmaceutical composition comprises one or more analgesic agents selected from the group consisting of aspirin, ibuprofen, naproxen, naproxen sodium, indomethacin, nabumetone and acetaminophen in an amount of 50-400 mg for each agent, and one or more antidiuretic agents, selected from the group consisting of antidiuretic hormone (ADH), angiotensin II, aldosterone, vasopressin, vasopressin analogs (e.g. desmopressin, argipressin, lipressin, felipressin, ornipressin, t rlipressina); vasopressin receptor agonists, atrial natriuretic peptide (ANP) receptor antagonists, and C-type natriuretic peptide (CNP) (i.e., NPR1, NPR2, NPR3) (e.g., HS-142-1, isatin [Asu7.23 '] b-ANP- (7-28)], anantine, a cyclic peptide from Streptomyces coerulescens and monoclonal antibody 3G12); type 2 somatostatin receptor antagonists (e.g., somatostatin), as well as their pharmaceutically acceptable derivatives, analogues, salts, hydrates and solvates, the pharmaceutical composition being formulated for sustained release with a two-phase release profile in which 20-60% of the active ingredients are released during 2 hours after administration and the remainder of the active ingredients is released continuously or at a constant rate for 5-24 hours, 5-8 hours, 8-16 hours or 16-24 hours.

In some embodiments, the pharmaceutical composition comprises one or more analgesic agents selected from the group consisting of aspirin, ibuprofen, naproxen, naproxen sodium, indomethacin, nabumetone and acetaminophen in an amount of 50-400 mg for each agent, and one or more antispasmodics selected from the group consisting of carisoprodol, benzodiazepines, baclofen, cyclobenzaprine, metaxalone, methocarbamol, clonidine, an analogue of clonidine and dantrolene in a total amount of 50-500 mg, while the pharmaceutical composition It is formulated for sustained release with a biphasic release profile in which 20-60% of the active ingredients are released within 2 hours after administration and the remainder of the active ingredients is released continuously or at a constant rate for 5-24 hours, 5-8 hours, 8-16 hours or 16-24 hours.

The present invention is further illustrated by the following examples, which should not be construed as limiting. The contents of all literature, patents and published patent applications mentioned in this application are incorporated herein by reference.

EXAMPLE 1: Suppression of urination to urinate

The study involved twenty volunteers, both men and women, each of whom experienced premature urges or a desire to urinate, which prevented them from sleeping for a sufficient period of time to feel well rested. Each subject took 400-800 mg of ibuprofen in a single dose at bedtime. At least 14 subjects reported that they could have a better rest, as they did not wake up as often as before due to the urge to urinate.

Some subjects reported that after several weeks of taking ibuprofen at night, the positive effect of less frequent urination disappeared. However, all these subjects then reported that the beneficial effect returned after several days of abstinence from taking doses.

EXAMPLE 2: EFFECT OF ANALGETIC MEDICINES, BOTULINIC NEUROTOXIN AND ANTIMUSKARIN MEDICINES ON THE RESPONSE RESPONSE OF MACROPHAGES TO INFLAMMATORY AND NON-INFLAMMATORY INCENTIVES

Experiment design

This study is intended to determine the dose and in vitro efficacy of analgesics and antimuscarinic agents in controlling the response of macrophages to inflammatory and non-inflammatory stimuli mediated by COX2 and prostaglandins (PGE, PGH, etc.). It sets the basic indicators (dose and kinetics) of responses to inflammatory and non-inflammatory effectors in the cells of the bladder. Briefly, cultured cells are exposed to analgesic and / or antimuscarinic agents in the absence or presence of various effectors.

Effectors include: lipopolysaccharide (LPS), an inflammatory agent, and a Cox2 inducer as an inflammatory stimulus; carbachol or acetylcholine, a stimulator of smooth muscle contraction, as a non-inflammatory stimulus; botulinum neurotoxin A, a known acetylcholine release inhibitor as a positive control, and arachidonic acid (AA), gamma-linolenic acid (DGLA) or eicosapentaenoic acid (EPA) as prostaglandin precursors that are produced after sequential oxidation of AA, DGLA or EPA cyclases inside the cell (COX1 and COX2) and end prostaglandin synthases.

Analgesic agents include: salicylates, such as aspirin, a derivative of isobutylphenylpropionic acid (ibuprofen), for example, advil, motrin, nuprin and mediprene, naproxen sodium, for example, olive, anaprox, antalgin, feminax ultra, flanax, inza, long-acting midol, nalgesin , naposin, naprozelin, naprogesic, naprosin, naprosin suspension, EC naprosin, narocin, proxene, synflex and xenobide, an acetic acid derivative such as indomethacin (indocin), a 1-naphthalene acetic acid derivative such as nabumetone or relafen, N-acetyl-para-aminophenol (APAP) bottoms, for example acetaminophen or paracetamol (tylenol), and celecoxib.

Antimuscarinic agents include: oxybutynin, solifenacin, darifenacin and atropine.

Macrophages are subjected to short-term (1-2 hours) or long-term (24-48 hours) stimulation using:

1) each analgesic agent individually in different doses,

(2) each analgesic in different doses in the presence of LPS,

(3) each analgesic in different doses in the presence of carbachol or acetylcholine,

(4) each analgesic in different doses in the presence of AA, DGLA or EPA,

(5) botulinum neurotoxin A alone at different doses,

(6) botulinum neurotoxin A at various doses in the presence of LPS,

(7) botulinum neurotoxin A in various doses in the presence of carbachol or acetylcholine,

(8) botulinum neurotoxin A in various doses in the presence of AA, DGLA or EPA,

(9) each antimuscarinic agent individually in different doses,

(10) each antimuscarinic agent in different doses in the presence of LPS,

(11) each antimuscarinic agent in different doses in the presence of carbachol or acetylcholine,

(12) each antimuscarinic agent in different doses in the presence of AA, DGLA or EPA.

Cells are then analyzed for the release of PGH2, PGE, PGE2, prostacidin, thromboxane, IL-1β, IL-6, TNF-α, COX2 activity, cAMP and cGMP production, IL-1β, IL-6, TNF-α and COX2 mRNA production as well as surface expression of the molecules of CD80, CD86 and MHC class II.

Materials and methods

Macrophages

In this study, murine macrophages of the RAW264.7 or J774 lines (obtained from ATCC) were used. Cells were maintained in culture medium containing RPMI 1640 supplemented with 10% fetal bovine serum (EBS), 15 mM HEPES, 2 mM L-glutamine, 100 U / ml penicillin and 100 μg / ml streptomycin. Cells were cultured at 37 ° C in an atmosphere of 5% CO ₂ and separated (reseeded) once a week.

In vitro analgesic treatment of macrophages

Macrophages of the RAW264.7 line were seeded in 96-well plates at a cell density of 1.5 × 10 ⁵ cells per well in 100 μl of culture medium. The cells were treated with (1) an analgesic (acetaminophen, aspirin, ibuprofen or naproxen) in various concentrations, (2) lipopolysaccharide (LPS), which is an effector of inflammatory stimuli for macrophages, in various concentrations, (3) carbachol or acetylcholine, which are non-inflammatory effectors stimuli, in various concentrations, (4) analgesic and LPS or (5) analgesic and carbachol or acetylcholine. Briefly, the analgesics were dissolved in an EBS-free culture medium (i.e., RPMI 1640 supplemented with 15 mM HEPES, 2 mM L-glutamine, 100 U / ml penicillin and 100 μg / ml streptomycin) and diluted to the desired concentrations by serial dilution in the same medium. For cells treated with an analgesic in the absence of LPS, 50 μl of analgesic solution and 50 μl of culture medium without EBS were added to each well. For cells treated with an analgesic in the presence of LPS, 50 μl of an analgesic solution and 50 μl of LPS (from Salmonella typhimurium) in a culture medium without EBS were added to each well. All processing conditions were tested in duplicate.

After 24 or 48 hours of cultivation, 150 μl of culture supernatants were collected, precipitated by centrifugation for 2 min at a speed of 8000 rpm at 4 ° C to remove cells and detritus and stored at -70 ° C for analysis of cytokine response by ELISA. Cells were collected and washed by centrifugation (5 min at a speed of 1500 rpm at 4 ° C) in 500 μl of phosphate buffer (PBS). Then half of the cells were quickly frozen in liquid nitrogen and stored at -70 ° C. The remaining cells were stained with fluorescently labeled monoclonal antibodies and analyzed by flow cytometry.

Flow cytometry analysis of expression of costimulatory molecules

For flow cytometric analysis, macrophages were diluted in 100 μl of FACS buffer (phosphate buffered saline (PBS) with 2% bovine serum albumin (BSA) and 0.01% NaN ₃ ) and stained for 30 min at 4 ° C by adding FITC-conjugated anti-CD40 antibodies, PE-conjugated anti-CD80 antibodies, PE-conjugated anti-CD86 antibodies, PE-conjugated anti-MHC class II (IA ^d ) antibodies (BD Bioscience). Then the cells were washed by centrifugation (5 min at a speed of 1500 rpm at 4 ° C) in 300 μl of FACS buffer. After a second wash, the cells were resuspended in 200 μl FACS buffer and the percentage of cells expressing a particular marker (positive) or a combination of markers (twice positive) was analyzed using an Accuri C6 flow cytometer (BD Biosciences).

Analysis of cytokine responses by ELISA

Cultural supernatants were analyzed by cytokine-specific ELISA to determine IL-1β, IL-6 and TNF-α responses in macrophage cultures treated only with analgesic, LPS, or a combination of LPS and analgesic. Nunc MaxiSorp immunological plates (Nunc) coated overnight with 100 μl mAbs anti-IL-6, mouse TNF-α (BD Biosciences) or anti-mouse IL-1β mAbs (R&D Systems) in 0.1 M sodium bicarbonate buffer were used for assays. (pH 9.5). After two washes with PBS (200 μl per well), 200 μl of PBS with 3% BSA was added to each well (block) and the plates were incubated for 2 hours at room temperature. The plates were again washed twice by adding 200 μl per well, then 100 μl of cytokine standards and culture supernatants were added in serial dilutions in duplicate, and the plates were incubated overnight at 4 ° C. Ultimately, the plates were washed twice and incubated with 100 μl of secondary biotinylated mAbs against IL-6, TNF-α (BD Biosciences) or IL-1β (R&D Systems) mice, followed by use of goat peroxidase-labeled mAbs against biotin (Vector Laboratories). A colorimetric reaction developed with the addition of a substrate of 2,2'-azino-bis (3) ethylbenzthiazolin-6-sulfonic acid (ABTS) and H ₂ O ₂ (Sigma) and the optical density was measured at 415 nm using a Victor ^® multi-plate reader V (PerkinElmer).

Determination of COX2 activity and cAMP and cGMP production

The activity of COX2 in cultured macrophages was determined by sequential competitive ELISA (R&D Systems). The production of cAMP and cGMP was determined using cAMP analysis and cGMP analysis. Such assays are common in the art.

results

Table 1 shows the results of experiments conducted with the Raw 264 macrophage cell line and the main results regarding the effect of analgesics on the expression of CD40 and CD80 costimulatory molecules on the cell surface. The expression of these molecules is stimulated by COX2 and inflammatory signals and, thus, evaluated in order to determine the functional consequences of COX2 inhibition.

As can be seen from table 2, acetaminophen, aspirin, ibuprofen and naproxen inhibit the basal expression of co-stimulatory molecules CD40 and CD80 by macrophages at all doses tested (i.e. 5 × 10 ⁵ nM, 5 × 10 ⁴ nM, 5 × 10 ³ nM, 5 × 10 ² nM, 50 nM and 5 nM), with the exception of the highest dose (i.e. 5 × 10 ⁶ nM), which seems to enhance and not inhibit the expression of co-stimulatory molecules. As shown in FIG. 1A and 1B, such an inhibitory effect on the expression of CD40 and CD50 was observed at such low doses of analgesics as 0.05 nM (i.e., 0.00005 μM). These results support the notion that controlled release of small doses of analgesics may be preferable to short-term delivery of high doses. The experiment also revealed that acetaminophen, aspirin, ibuprofen, and naproxen have a similar inhibitory effect on LPS-induced expression of CD40 and CD80.

Table 1 Experiment design The control LPS
Salmonella typhimurium Acetaminophen Aspirin Ibuprofen Naproxen TESTS one X 2 X Dose response (0, 5, 50, 1000) ng / ml 3 X Dose responses (0, 5, 50, 500, 5 × 10 ³ , 5 × 10 ⁴ , 5 × 10 ⁵ , 5 × 10 ⁶ ) nM four X X (5 ng / ml) X (50 ng / ml X (1000 ng / ml) Dose responses (0, 5, 50, 500, 5 × 10 ³ , 5 × 10 ⁴ , 5 × 10 ⁵ , 5 × 10 ⁶ ) nM

ANALYSIS a Activation / Stimulation Status Characterization: Flow Cytometry Assay CD40, CD80, CD86 and MHC Class II b Inflammatory response mediators: analysis of IL-1β, IL-6, TNF-α by ELISA

table 2 Summary main experimental results Effectors % positive Negative control LPS 5 ng / ml Analgesic dose
(nM) 5 × 10 ⁶ 5 × 10 ⁵ 5 × 10 ⁴ 5 × 10 ³ 500 fifty 5 CD40 ⁺ CD80 ⁺ 20.6 77.8 Acetaminophen CD40 ⁺ CD80 ⁺ 63 eighteen 12 9.8 8.3 9.5 7.5 Aspirin CD40 ⁺ CD80 ⁺ 44 eleven 10.3 8.3 8 10.5 7.5 Ibuprofen CD40 ⁺ CD80 ⁺ HB * 6.4 7.7 7.9 6.0 4.9 5.8 Naproxen CD40 ⁺ CD80 ⁺ 37 9.6 7.7 6.9 7.2 6.8 5.2 Analgesic plus LPS Acetaminophen CD40 ⁺ CD80 ⁺ 95.1 82.7 72,4 68.8 66.8 66,2 62.1 Aspirin CD40 ⁺ CD80 ⁺ 84.5 80 78.7 74.7 75.8 70.1 65.7 Ibuprofen CD40 ⁺ CD80 ⁺ HB 67 77.9 72.9 71.1 63.7 60.3 Naproxen CD40 ⁺ CD80 ⁺ 66.0 74.1 77.1 71.0 68.8 72 73 * HB: not fulfilled (toxicity)

Table 3 summarizes the results of several studies in which serum levels of analgesics were measured after oral administration of therapeutic doses to adults. As can be seen from table 3, the maximum serum levels of analgesics after oral administration of therapeutic doses are in the range from 10 ⁴ to 10 ⁵ nm. Thus, the in vitro tested doses of the analgesics shown in Table 2 cover the range of concentrations achieved in vivo in humans.

Table 3 Levels of analgesics in human serum after oral administration of therapeutic doses Analgesic Molecular mass Maximum serum levels after oral administration of therapeutic doses References mg / l nM Acetaminophen (Tylenol) 151.16 11-18 7.2 × 10 ⁴ -1.19 × 10 ⁵ * BMC Clinical Pharmacology, 2010, 10:10
* Anaesth Intensive Care, 2011, 39: 242 Aspirin (Acetylsalicylic acid) 181.66 30-100 1.65 × 10 ⁵ -5.5 × 10 ⁵ * Disposition of Toxic Drugs and Chemicals in Man, 8th Edition, Biomedical Public, Foster City, CA, 2008, pp, 22-25
* J Lab Clin Med, 1984 Jun; 103: 869 Ibuprofen (Advil, Motrin) 206.29 24-32 1.16 × 10 ⁵ -1.55 × 10 ⁵ * BMC Clinical Pharmacology2010, 10:10 * J Clin Pharmacol, 2001, 41: 330 Naproxen (Olive) 230.26 up to 60 up to 2.6 × 10 ⁵ * J Clin Pharmacol, 2001, 41: 330

EXAMPLE 3: EFFECT OF ANALGETIC MEDICINES, BOTULINIC NEUROTOXIN AND ANTIMUSKARIN MEDICINES ON THE RESPONSE OF MUSCLE BLADDERS OF THE BLADDER TO AN INFLAMMATORY AND NON-INFLAMMATORY

Experiment design

This study is intended to determine how optimal doses of the analgesics defined in Example 2 affect smooth muscle cells of the bladder in cell culture or tissue cultures, and to determine whether various classes of analgesics can act synergistically to more effectively inhibit COX2 and PGE2 responses. .

Effectors, analgesic agents, and antimuscarinic agents are described in Example 2.

The primary culture of smooth muscle cells of the mouse bladder is subjected to short-term (1-2 hours) or long-term (24-48 hours) stimulation using:

1) each analgesic agent individually in different doses,

(2) each analgesic in different doses in the presence of LPS,

(3) each analgesic in different doses in the presence of carbachol or acetylcholine,

(4) each analgesic in different doses in the presence of AA, DGLA or EPA,

(5) botulinum neurotoxin A alone at different doses,

(6) botulinum neurotoxin A at various doses in the presence of LPS,

(7) botulinum neurotoxin A in various doses in the presence of carbachol or acetylcholine,

(8) botulinum neurotoxin A in various doses in the presence of AA, DGLA or EPA,

(9) each antimuscarinic agent individually in different doses,

(10) each antimuscarinic agent in different doses in the presence of LPS,

(11) each antimuscarinic agent in different doses in the presence of carbachol or acetylcholine,

(12) each antimuscarinic agent in different doses in the presence of AA, DGLA or EPA.

Cells are then analyzed for the release of PGH2, PGE, PGE2, prostacidin, thromboxane, IL-1β, IL-6, TNF-α, COX2 activity, cAMP and cGMP production, IL-1β, IL-6, TNF-α and COX2 mRNA production as well as surface expression of the molecules of CD80, CD86 and MHC class II.

Materials and methods

Isolation and purification of mouse bladder cells

The bladder was removed from euthanized C57BL / 6 mice (8-12 weeks old) and the cells were isolated by enzymatic digestion, followed by purification on a percoll gradient. Briefly, bladders from 10 mice were sheared with scissors to a thin suspension in 10 ml splitting buffer (RPMI 1640, 2% bovine fetal serum, 0.5 mg / ml collagenase, 30 μg / ml DNase). Bladder suspensions were enzymatically digested for 30 minutes at 37 ° C. Unsplit fragments were further dispersed using a Cell-tainer ^® device. A cell suspension was besieged and added to a stepwise gradient of 20%, 40% and 75% percoll for purification of mononuclear cells. In each experiment, 50-60 bladders were used.

After washing in RPMI 1640, bladder cells were resuspended in RPMI 1640 supplemented with 10% fetal bovine serum, 15 mM HEPES, 2 mM L-glutamine, 100 U / ml penicillin and 100 μg / ml streptomycin, and plated in 96-black transparent bottom microculture well plates at a cell density of 3 × 10 ⁴ cells per well in a volume of 100 μl. Cells were cultured at 37 ° C in an atmosphere of 5% CO ₂ .

In vitro analgesic treatment of cells

Bladder cells were treated with analgesic solutions (50 μl / well) either individually or in combination with carbachol (10 molar, 50 μl / well) as an example of a non-inflammatory stimulus, or lipopolysaccharide (LPS) from Salmonella typhimurium (1 μg / ml, 50 μl / well), as an example of an inflammatory stimulus. If no other effectors were added to the cells, 50 μl of RPMI 1640 without fetal bovine serum was added to the wells to bring the final volume to 200 μl.

After 24 hours of cultivation, 150 μl of culture supernatants were collected, centrifuged for 2 min at a speed of 8000 rpm at 4 ° C to remove cells and detritus and stored at -70 ° C for analysis of prostaglandin E2 (PGE2) ELISA responses. Cells were fixed, permeabilized and blocked to detect cyclooxygenase-2 (COX2) using a fluorogenic substrate. In a specific experiment, cells were stimulated 12 hours in vitro to analyze COX2 responses.

Analysis of COX2 responses

COX2 responses were analyzed by cell ELISA using the human / mouse total COX2 immunoassay kit (R&D Systems) following the manufacturer's instructions. Shortly after cell fixation and permeabilization, mouse antibodies against all COX2 and rabbit antibodies against total GAPDH were added to the wells of black with a clear bottom 96-well microculture plates. After incubation and washing, PH-conjugated anti-mouse IgG antibodies and alkaline phosphatase-conjugated anti-rabbit IgG antibodies were added to the wells. After the next incubation and a series of washes, fluorogenic substrates for HRP and alkaline phosphatase were added. Ultimately, a Victor ^® V multiple-label plate reader was used to record the fluorescence emitted at a wavelength of 600 nm (COX2 fluorescence) and 450 nm (GAPDH fluorescence). The results are presented as relative levels of all COX2, expressed in relative fluorescence units (OEF) (RFUs) and normalized to the “household” GAPDH protein.

Analysis of PGE2 responses

Prostaglandin E2 responses were analyzed by sequential competitive ELISA (R&D Systems). In more detail, culture supernatants or PGE2 standards were added to the wells of a 96-well microtiter microtiter plate coated with goat polyclonal antibodies against mouse immunoglobulins. After an hour of incubation on a plate shaker, PC-conjugated PGE2 was added and the plates were incubated for another two hours at room temperature. The plates were then washed and a HRP substrate solution was added to each well. The color developed for 30 minutes and the reaction was stopped by the addition of sulfuric acid before reading in the wells of a plate at a wavelength of 450 nm with a correction of wavelength at 570 nm. Results are expressed as mean pg / ml PGE2.

Other analyzes

Release of PGH2, PGE, prostacidin, thromboxane, IL-1β, IL-6 and TNF-α, cAMP and cGMP production, mRNA production of IL-1β, IL-6, TNF-α and COX2, as well as surface expression of CD80, CD86 molecules and MHC class II was determined as described in example 2.

results

Analgesics inhibit mouse bladder COX2 responses to inflammatory stimuli

Several analgesics (acetaminophen, aspirin, ibuprofen, and naproxen) were tested on mouse bladder cells at a concentration of 5 μM or 50 μM to determine whether analgesics can induce COX2 responses. Analysis of 24-hour cultures showed that none of the tested analgesics induced COX2 responses in mouse urinary bladder cells in vitro.

The effect of these analgesics on COX2 responses of bladder cells to stimulation with carbachol or LPS in vitro was also tested. As indicated in Table 1, carbachol in the tested dose did not have a significant effect on COX2 levels in mouse bladder cells. LPS, on the other hand, significantly increased overall COX2 levels. Interestingly, acetaminophen, aspirin, ibuprofen, and naproxen all could suppress the effect of LPS on COX2 levels. The inhibitory effect of analgesics was noticeable when these drugs were tested at either 5 μM or 50 μM (Table 4).

Table 4 Expression of COX2 by mouse bladder cells after in vitro stimulation and analgesic treatment Stimulus Analgesic General COX2 levels
(normalized OEF) no no 158 ± 18 Carbachol (mm) no 149 ± 21 LPS (1 μg / ml) no 420 ± 26 LPS (1 μg / ml) Acetaminophen (5 μM) 275 ± 12 LPS (1 μg / ml) Aspirin (5 μM) 240 ± 17 LPS (1 μg / ml) Ibuprofen (5 μM)) 253 ± 32 LPS (1 μg / ml) Naproxen (5 μM) 284 ± 11 LPS (1 μg / ml) Acetaminophen (50 μM) 243 ± 15 LPS (1 μg / ml) Aspirin (50 μM) 258 ± 21 LPS (1 μg / ml) Ibuprofen (50 μM) 266 ± 19 LPS (1 μg / ml) Naproxen (50 μM) 279 ± 23

Analgesics inhibit PGE2 responses of mouse bladder cells to inflammatory stimulus

The secretion of PGE2 in the culture supernatants of mouse bladder cells was measured to determine the biological significance of analgesic changes in COX2 levels in mouse bladder cells. As shown in table 5, PGE2 was not found in the culture supernatants of unstimulated bladder cells or bladder cells cultured in the presence of carbachol. As with the COX2 responses described above, stimulation of the mouse LPS bladder cells induced the secretion of large amounts of PGE2. The addition of acetaminophen, aspirin, ibuprofen, and naproxen analgesics suppressed the effect of LPS on PGE2 secretion, and no difference was found between the responses of cells treated with analgesics at a dose of 5 or 50 μM.

Table 5 The secretion of PGE2 by mouse bladder cells after in vitro stimulation and analgesic treatment Stimulus Analgesic PGE2 levels (pg / ml) no no <20.5 Carbachol (mm) no <20.5 LPS (1 μg / ml) no 925 ± 55 LPS (1 μg / ml) Acetaminophen (5 μM) 619 ± 32 LPS (1 μg / ml) Aspirin (5 μM) 588 ± 21 LPS (1 μg / ml) Ibuprofen (5 μM)) 593 ± 46 LPS (1 μg / ml) Naproxen (5 μM) 597 ± 19 LPS (1 μg / ml) Acetaminophen (50 μM) 600 ± 45 LPS (1 μg / ml) Aspirin (50 μM) 571 ± 53 LPS (1 μg / ml) Ibuprofen (50 μM) 568 ± 32 LPS (1 μg / ml) Naproxen (50 μM) 588 ± 37

Thus, these data show that analgesics alone at a concentration of 5 μM or 50 μM do not induce COX2 and PGE2 responses in mouse bladder cells. However, analgesics at a dose of 5 μM or 50 μM significantly inhibit the COX2 and PGE2 responses of mouse urinary bladder cells stimulated by in vitro LPS (1 μg / ml). No significant effect of the analgesics on COX2 and PGE2 was observed for carbachol-stimulated urinary bladder cells (1 mM).

EXAMPLE 4: EFFECT OF ANALGETIC MEDICINES, BOTULINIC NEUROTOXIN AND ANTIMUSCARINE MEDICINES ON THE REDUCTION OF MUSCLE BLADDERS OF THE BLADDER OF THE MOUSE

Experiment design

Cultured smooth muscle cells of a mouse or rat bladder and smooth muscle tissue of a mouse or rat bladder are exposed to inflammatory stimuli and non-inflammatory stimuli in the presence of an analgesic agent and / or antimuscarinic agent in various concentrations. Stimulus-induced contraction is measured to evaluate the inhibitory effect of the analgesic and / or antimuscarinic.

Effectors, analgesic agents, and antimuscarinic agents are described in Example 2.

The primary culture of smooth muscle cells of the mouse bladder is subjected to short-term (1-2 hours) or long-term (24-48 hours) stimulation using:

1) each analgesic agent individually in different doses,

(2) each analgesic in different doses in the presence of LPS,

(3) each analgesic in different doses in the presence of carbachol or acetylcholine,

(4) each analgesic in different doses in the presence of AA, DGLA or EPA,

(5) botulinum neurotoxin A alone at different doses,

(6) botulinum neurotoxin A at various doses in the presence of LPS,

(7) botulinum neurotoxin A in various doses in the presence of carbachol or acetylcholine,

(8) botulinum neurotoxin A in various doses in the presence of AA, DGLA or EPA,

(9) each antimuscarinic agent individually in different doses,

(10) each antimuscarinic agent in different doses in the presence of LPS,

(11) each antimuscarinic agent in different doses in the presence of carbachol or acetylcholine,

(12) each antimuscarinic agent in different doses in the presence of AA, DGLA or EPA.

Materials and methods

Primary mouse bladder cells were isolated as described in Example 3. Bladder tissue cultures were used in certain experiments. Bladder smooth muscle contractions were recorded on a Grass polygraph (Quincy Mass, USA).

EXAMPLE 5: EFFECT OF ORAL ANALGETIC AGENTS AND ANTIMUSKARIN MEDICINES ON COX2 AND PGE2 MUSCLE BLADDER RESPONSES OF THE MOUSE

Experiment design

Oral doses of aspirin, sodium naproxen, ibuprofen, indocin, nabumeton, tylenol, celecoxib, oxybutynin, solifenacin, darifenacin, atropine, and combinations thereof are orally administered to normal mice and mice with overactive bladder syndrome. Control groups include drug-free normal mice and drug-free mice with Overactive Bladder Syndrome (OAB). Thirty (30) minutes after the last doses, the bladders are collected and stimulated ex vivo with carbachol or acetylcholine. In certain experiments, the bladders are treated with botulinum neurotoxin A before stimulation with carbachol. Animals are kept in metabolic cells and the frequency (and volume) of urination is assessed. Bladder productivity is determined by monitoring water intake and litter weight in cells. Serum levels of PGH2, PGE, PGE2, prostacidin, thromboxane, IL-1β, IL-6, TNF-α, cAMP and cGMP are determined by ELISA. The expression of CD80, CD86, MHC class II in whole blood cells is determined by flow cytometry.

At the end of the experiment, the animals are euthanized and bladder contractions ex vivo are recorded on a Grass polygraph. Parts of the bladder are fixed in formalin and COX2 responses are analyzed by immunohistochemical methods.

EXAMPLE 6: EFFECT OF ANALGETIC MEDICINES, BOTULINIC NEUROTOXIN AND ANTIMUSKARIN MEDICINES ON THE REACTION OF HUMAN BLADDERS IN BLINDERS AND INFLAMMATIS

Experiment design

This study is intended to determine how the optimal doses of the analgesics defined in Examples 1-5 affect smooth muscle cells in the human bladder in cell culture or tissue cultures, and to determine whether analgesics of different classes can act synergistically to more effectively inhibit COX2 and PGE2 responses.

Effectors, analgesic agents, and antimuscarinic agents are described in Example 2.

Smooth muscle cells of the human bladder are subjected to short-term (1-2 hours) or long-term (24-48 hours) stimulation using:

1) each analgesic agent individually in different doses,

(2) each analgesic in different doses in the presence of LPS,

(3) each analgesic in different doses in the presence of carbachol or acetylcholine,

(4) each analgesic in different doses in the presence of AA, DGLA or EPA,

(5) botulinum neurotoxin A alone at different doses,

(6) botulinum neurotoxin A at various doses in the presence of LPS,

(7) botulinum neurotoxin A in various doses in the presence of carbachol or acetylcholine,

(8) botulinum neurotoxin A in various doses in the presence of AA, DGLA or EPA,

(9) each antimuscarinic agent individually in different doses,

(10) each antimuscarinic agent in different doses in the presence of LPS,

(11) each antimuscarinic agent in different doses in the presence of carbachol or acetylcholine,

(12) each antimuscarinic agent in different doses in the presence of AA, DGLA or EPA.

Cells are then analyzed for the release of PGH2, PGE, PGE2, prostacidin, thromboxane, IL-1β, IL-6, TNF-α, COX2 activity, cAMP and cGMP production, IL-1β, IL-6, TNF-α and COX2 mRNA production as well as surface expression of the molecules of CD80, CD86 and MHC class II.

EXAMPLE 7: EFFECT OF ANALGETIC MEDICINES, BOTULINIC NEUROTOXIN AND ANTIMUSKARIN MEDICINES ON THE REDUCTION OF HUMAN BLADDERS

Experiment design

Cultivated smooth muscle cells of the human bladder are exposed to inflammatory stimuli and non-inflammatory stimuli in the presence of an analgesic and / or antimuscarinic in various concentrations. Stimulus-induced contraction is measured to evaluate the inhibitory effect of the analgesic and / or antimuscarinic.

Effectors, analgesic agents, and antimuscarinic agents are described in Example 2.

Smooth muscle cells of the human bladder are subjected to short-term (1-2 hours) or long-term (24-48 hours) stimulation using:

1) each analgesic agent individually in different doses,

(2) each analgesic in different doses in the presence of LPS,

(3) each analgesic in different doses in the presence of carbachol or acetylcholine,

(4) each analgesic in different doses in the presence of AA, DGLA or EPA,

(5) botulinum neurotoxin A alone at different doses,

(6) botulinum neurotoxin A at various doses in the presence of LPS,

(7) botulinum neurotoxin A in various doses in the presence of carbachol or acetylcholine,

(8) botulinum neurotoxin A in various doses in the presence of AA, DGLA or EPA,

(9) each antimuscarinic agent individually in different doses,

(10) each antimuscarinic agent in different doses in the presence of LPS,

(11) each antimuscarinic agent in different doses in the presence of carbachol or acetylcholine,

(12) each antimuscarinic agent in different doses in the presence of AA, DGLA or EPA.

Bladder smooth muscle contractions are recorded on a Grass polygraph (Quincy Mass, USA).

EXAMPLE 8: ANALGETIC EFFECTS ON THE RESPONSE OF HUMAN NORMAL BLADDER CELLS TO THE INFLAMMATORY AND NON-INFLAMMATORY SIGNALS

Experiment design

Culture of smooth muscle cells of a normal human bladder

Smooth muscle cells of a normal human bladder were isolated by enzymatic digestion of macroscopically normal human bladder fragments. Cells were grown in vitro by culturing at 37 ° C in an atmosphere of 5% CO ₂ in RPMI 1640 medium supplemented with 10% fetal bovine serum, 15 mM HEPES, 2 mM L-glutamine, 100 U / ml penicillin and 100 mg / ml streptomycin and reseeded once a week, treated with trypsin to detach cells, followed by reseeding in a new culture mattress. In the first week of cultivation, epidermal growth factor at a concentration of 0.5 ng / ml, fibroblast growth factor at a concentration of 2 ng / ml and insulin at a concentration of 5 μg / ml were added to the culture medium.

Treatment of smooth muscle cells of a normal human bladder with in vitro analgesics

Bladder smooth muscle cells treated with trypsin and seeded in microculture plates at a cell density of 3 × 10 ⁴ cells per well in a volume of 100 μl were treated with analgesic solutions (50 μl / well) either individually or in combination with carbachol (10 molar) 50 μl / well), as an example of a non-inflammatory stimulus, or by lipopolysaccharide (LPS) from Salmonella typhimurium (1 μg / ml, 50 μl / well), as an example of an inflammatory stimulus. If no other effectors were added to the cells, 50 μl of RPMI 1640 without fetal bovine serum was added to the wells to bring the final volume to 200 μl.

After 24 hours of cultivation, 150 μl of culture supernatants were collected, centrifuged for 2 min at a speed of 8000 rpm at 4 ° C to remove cells and detritus and stored at -70 ° C for analysis of prostaglandin E2 (PGE2) ELISA responses. Cells were fixed, permeabilized and blocked to detect COX2 using a fluorogenic substrate. In a specific experiment, cells were stimulated 12 hours in vitro to analyze COX2, PGE2 and cytokine responses.

Analysis of COX2, PGE2 and cytokine responses

COX2 and PGE2 responses were analyzed as described in Example 3. Cytokine responses were analyzed as described in Example 2.

results

Analgesics inhibit COX2 responses of smooth muscle cells in a normal human bladder to inflammatory and non-inflammatory stimuli - Analysis of cells and culture supernatants after 24 hours of culture showed that none of the analgesics tested individually induced COX2 responses in smooth muscle cells of a normal human bladder. However, as shown in Table 6, carbachol induced low but significant COX2 responses in smooth muscle cells of a normal human bladder. On the other hand, LPS treatment led to higher levels of COX2 responses in the smooth muscle cells of a normal human bladder. Acetaminophen, aspirin, ibuprofen, and naproxen could all suppress the effect of carbochol and LPS on COX2 levels. The overwhelming effect of analgesics was noticeable with respect to LPS-induced responses when these drugs were tested at either 5 μM or 50 μM.

Table 6 Expression of COX2 by smooth muscle cells of a normal human bladder after in vitro stimulation with inflammatory and non-inflammatory stimuli and treatment with analgesics Stimulus Analgesic General COX2 levels ^#
(normalized OEF)
subject 1 General COX2 levels
(normalized OEF)
subject 2 no no 230 199 Carbachol 10 ^-3 M no (50 μM) 437 462 Carbachol 10 ^-3 M Acetaminophen (50 μM) 298 310

Carbachol 10 ^-3 M Aspirin (50 μM) 312 297 Carbachol 10 ^-3 M Ibuprofen (50 μM) 309 330 Carbachol 10 ^-3 M Naproxen (50 μM) 296 354 LPS (10 μg / ml) no 672 633 LPS (10 μg / ml) Acetaminophen (5 μM) 428 457 LPS (10 μg / ml) Aspirin (5 μM) 472 491 LPS (10 μg / ml) Ibuprofen (5 μM) 417 456 LPS (10 μg / ml) Naproxen (5 μM) 458 501 LPS (10 μg / ml) Acetaminophen (50 μM) 399 509 LPS (10 μg / ml) Aspirin (50 μM) 413 484 LPS (10 μg / ml) Ibuprofen (50 μM) 427 466 LPS (10 μg / ml) Naproxen (50 μM) 409 458 ^# Data are the average of two repetitions

Analgesics inhibit the PGE2 responses of smooth muscle cells of a normal human bladder to inflammatory and non-inflammatory stimuli - As with the induction of COX2 responses described above, both carbachol and LPS induced PGE2 production by smooth muscle cells of a normal human bladder. It was also found that acetaminophen, aspirin, ibuprofen, and naproxen suppress LPS-induced PGE2 responses in both 5 μM and 50 μM concentrations (Table 7).

Table 7 Secretion of PGE2 by smooth muscle cells of the normal human bladder after in vitro stimulation with inflammatory and non-inflammatory stimuli and analgesic treatment Stimulus Analgesic PGE2 ^# levels (pg / ml)
Subject 1 PGE2 levels (pg / ml)
Subject 2 no no <20.5 <20.5 Carbachol 10 ^-3 M no 129 104 Carbachol 10 ^-3 M Acetaminophen (50 μM) 76 62 Carbachol 10 ^-3 M Aspirin (50 μM) 89 59 Carbachol 10 ^-3 M Ibuprofen (50 μM) 84 73 Carbachol 10 ^-3 M Naproxen (50 μM) 77 66 LPS (10 μg / ml) no 1125 998 LPS (10 μg / ml) Acetaminophen (5 μM) 817 542 LPS (10 μg / ml) Aspirin (5 μM) 838 598 LPS (10 μg / ml) Ibuprofen (5 μM) 824 527 LPS (10 μg / ml) Naproxen (5 μM) 859 506 LPS (10 μg / ml) Acetaminophen (50 μM) 803 540 LPS (10 μg / ml) Aspirin (50 μM) 812 534 LPS (10 μg / ml) Ibuprofen (50 μM) 821 501 LPS (10 μg / ml) Naproxen (50 μM) 819 523 ^# Data are the average of two repetitions

Analgesics inhibit the cytokine responses of smooth muscle cells of the normal human bladder to inflammatory stimuli - Analysis of cells and culture supernatants after 24 hours of cultivation showed that none of the analgesics tested individually induced secretion of IL-6 or TNFα in the smooth muscle cells of normal human bladder . As shown in tables 8 and 9, carbachol in the tested doses induced low but significant TNFα and IL-6 responses in smooth muscle cells of a normal human bladder. On the other hand, treatment with LPS led to massive induction of these pro-inflammatory cytokines. Acetaminophen, aspirin, ibuprofen, and naproxen suppressed the effect of carbachol and LPS on TNFα and IL-6 responses. The overwhelming effect of analgesics was noticeable with respect to LPS-induced responses when these drugs were tested at either 5 μM or 50 μM.

Table 8 Secretion of TNFα by smooth muscle cells of the normal human bladder after in vitro stimulation with inflammatory and non-inflammatory stimuli and analgesic treatment Stimulus Analgesic TNFα (pg / ml) ^#
Subject 1 TNFα (pg / ml)
Subject 2 no no <5 <5 Carbachol 10 ^-3 M no 350 286 Carbachol 10 ^-3 M Acetaminophen (50 μM) 138 164 Carbachol 10 ^-3 M Aspirin (50 μM) 110 142 Carbachol 10 ^-3 M Ibuprofen (50 μM) 146 121 Carbachol 10 ^-3 M Naproxen (50 μM) 129 137 LPS (10 μg / ml) no 5725 4107 LPS (10 μg / ml) Acetaminophen (5 μM) 2338 2267 LPS (10 μg / ml) Aspirin (5 μM) 2479 2187 LPS (10 μg / ml) Ibuprofen (5 μM) 2733 2288 LPS (10 μg / ml) Naproxen (5 μM) 2591 2215 LPS (10 μg / ml) Acetaminophen (50 μM) 2184 2056

LPS (10 μg / ml) Aspirin (50 μM) 2266 2089 LPS (10 μg / ml) Ibuprofen (50 μM) 2603 1997 LPS (10 μg / ml) Naproxen (50 μM) 2427 2192 ^# Data are the average of two repetitions Table 9 Secretion of IL-6 by smooth muscle cells of a normal human bladder after in vitro stimulation with inflammatory and non-inflammatory stimuli and treatment with analgesics Stimulus Analgesic IL-6 (pg / ml) ^#
Subject 1 IL-6 (pg / ml)
Subject 2 no no <5 <5 Carbachol 10 ^-3 M no 232 278 Carbachol 10 ^-3 M Acetaminophen (50 μM) 119 135 Carbachol 10 ^-3 M Aspirin (50 μM) 95 146 Carbachol 10 ^-3 M Ibuprofen (50 μM) 107 118 Carbachol 10 ^-3 M Naproxen (50 μM) 114 127 LPS (10 μg / ml) no 4838 4383 LPS (10 μg / ml) Acetaminophen (5 μM) 2012 2308 LPS (10 μg / ml) Aspirin (5 μM) 2199 2089 LPS (10 μg / ml) Ibuprofen (5 μM) 2063 2173 LPS (10 μg / ml) Naproxen (5 μM) 2077 2229 LPS (10 μg / ml) Acetaminophen (50 μM) 2018 1983 LPS (10 μg / ml) Aspirin (50 μM) 1987 2010 LPS (10 μg / ml) Ibuprofen (50 μM) 2021 1991 LPS (10 μg / ml) Naproxen (50 μM) 2102 2028 ^# Data are the average of two repetitions

Primary smooth muscle cells of a normal human bladder were isolated, cultured, and evaluated for their responses to analgesics in the presence of non-inflammatory (carbachol) and inflammatory (LPS) stimuli. The purpose of this study was to determine whether the observations made previously for mouse bladder cells may or may not be applicable to smooth muscle cells of a normal human bladder.

The above experiment will be reproduced with analgesic agents and / or antimuscarinic agents in a delayed release or sustained release preparation or delayed and prolonged release preparations.

The above description is intended to explain to an ordinary person skilled in the art how to put the present invention into practice, and it is not intended to describe in detail all the obvious modifications and variations that will become apparent to a skilled person when reading this description. It is contemplated, however, that all such obvious modifications and variations will fall within the scope of the present invention, which is defined by the appended claims. The claims should encompass the claimed components and steps in any order that are effective for the achievement of the intended objectives, unless the context clearly indicates otherwise.

Claims (10)

1. A method of reducing the frequency of urination in a subject, including: administering to a subject in need of a pharmaceutical composition consisting of: effective amounts of zolpidem and one or more of the analgesic agents; and a pharmaceutically acceptable carrier wherein said one or more of the analgesic agents are formulated for sustained or delayed release and wherein said zolpidem is formulated for immediate release. 2. A pharmaceutical composition for reducing the frequency of urination in a subject, consisting of: effective amounts of zolpidem and one or more of the analgesic agents; and a pharmaceutically acceptable carrier wherein said one or more of the analgesic agents are formulated for sustained or delayed release and wherein said zolpidem is formulated for immediate release.

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