Patent Application
20150045392
The embodiments disclosed herein relate to methods of norketotifen-treatment of medical conditions that are mediated through histamine H-4 receptors in mammals.
Four histamine receptors (H-1, H-2, H-3, H-4) have been identified, all of which are G protein-coupled receptors. The four different receptors are expressed on various cell types and exert their effects through different intracellular signaling mechanisms, which may in part be related to the diverse effects of histamine in different cells, tissues and organs.
The human histamine H-4 receptor is active constitutively, as is also the case with human histamine H-1, H-2 and H-3 receptors. Thus, like all other histamine receptor inhibitors, histamine H-4 receptor inhibitors are inverse agonists on the human histamine receptors.
Several potent and selective histamine H-4 receptor ligands have been described, such as for example JNJ 7777120, JNJ 10191584 and 4-methylhistamine. Such ligands will bind to all human histamine H-4 receptors, wherever those receptors are expressed in the body, such as for example, in bone marrow and white blood cells. The disadvantage of these compounds is that they act with high potency inhibiting histamine H-4 receptors throughout the body, increasing the likelihood of unwanted systemic side effects.
What is needed are improved methods of selectively treating medical conditions associated with histamine H-4 receptors at the biophases of those conditions.
In one aspect, disclosed herein is a method of treating a mammal in need of treatment for histamine H-4 receptor-related pruritus, comprising orally administering to the mammal in need thereof a therapeutically effective amount of RS-norketotifen or a pharmaceutically acceptable salt thereof. In specific embodiments, administration of the therapeutically effective amount of norketotifen or a pharmaceutically acceptable salt thereof is expected to avoid systemic histamine-H-4-related adverse drug effects, because the orally administered amount of the drug accumulates in the skin, which is the largest organ in the body.
In another aspect, a method of treating a mammal in need of treatment for histamine H-4 receptor-related pruritus without inducing systemic side effects is described and comprises orally administering to the mammal in need thereof a therapeutically effective amount of a biophase-selective histamine H-4 receptor inhibitor, wherein the biophase-selective histamine H-4 receptor inhibitor is norketotifen, an isomer of norketotifen, or a pharmaceutically acceptable salt of racemic or isomeric norketotifen. Pruritus (itch) is a sensation that causes the desire or reflex to scratch. Pain and pruritus have anatomical and physiological similarities, but while pain evokes a withdrawal reflex, pruritus creates a scratching reflex. Often multiple scratches are evoked, usually called “bouts of scratches”. The word “pruritus” is Latin for “itch”.
Histamine H-4 receptors are known to be expressed in the skin as well as in the pulmonary and gastrointestinal tissues of humans and of animals. Histamine H-4 receptors are also expressed in bone marrow, and serious side effects, such as for example, agranulocytosis, are believed to be related to the activity of potent histamine H-4 receptor inhibitors. Without being held to theory, it is believed that systemic adverse effects will be less pronounced for compounds that are selectively distributed to specific biophase organs or tissues where the compounds may accumulate in concentrations that are higher than the systemic plasma concentration. Such compounds will reach concentrations that inhibit histamine H-4 receptors locally in the specific organs or tissues, thereby reducing the systemic exposure and the incidence of systemic side effects. Thus, rather than increasing the potency or the dose of histamine H-4 receptor ligands, it may be advantageous to use a histamine H-4 receptor inhibitor that accumulates at the biophase(s) for diseases. Such compounds will selectively express histamine H-4 receptor inverse agonistic activity where they are accumulated, while avoiding the adverse effects of evenly distributed ligands with high affinity for the histamine H-4 receptors.
As used herein, the term biophase refers to the site(s) where a drug expresses its therapeutic activity. For example, if a receptor is associated with a disorder in the skin, then the biophase is the skin.
As used herein, a potent histamine H-4 receptor inverse agonist has a Ki less than 3.0E-7M, while moderately active compounds have Ki between 3.0E-6M and 3.0E-7M.
It is highly advantageous if compounds with moderate histamine H-4 receptor activity accumulate where they are anatomically therapeutically active; the moderate activity will not express systemic adverse effects, such as for example agranulocytosis. Such compounds are herein called biophase-selective histamine H-4 receptor ligands or biophase-selective histamine H-4 inhibitors or biophase-selective histamine H-4 inverse agonists.
It has specifically been found that after oral administration, racemic norketotifen (RS-norketotifen) and the isomers thereof (R-norketotifen, S-norketotifen) accumulate in the skin. The skin is the biophase for dermal diseases and the pharmacokinetic exposure parameters—area under the curve, half-lives and mean residence times—of norketotifen in this biophase have been found to significantly exceed the corresponding parameters in the systemic circulation (See, e.g., Example 4.) The high concentrations of these compounds in skin (See, e.g., Example 4) will inhibit the histaminic H-4 receptors (See, e.g., Examples 2 and 3), particularly since these compounds potently decrease the concentrations of available histamine at the biophase (See, e.g., Example 1). Thus, RS-, R- and S-norketotifen, as well as other histamine H-4 receptor inhibitors that are distributed in high concentrations in the skin, can advantageously be used to treat diseases or medical conditions associated with dermal histamine H-4 receptors.
Further, norketotifen and the isomers thereof have moderate affinity for histamine H-4 receptors. This moderate affinity combined with the localization in the biophases of the skin and lungs avoids adverse effects induced by systemically active histamine H-4 receptor inhibitors.
In one embodiment, disclosed herein is a method treating a mammal in need of treatment for histamine H-4 receptor-related pruritus, comprising orally administering to the mammal in need thereof a therapeutically effective amount of a biophase-specific H-4-receptor inhibitor. In specific embodiments, the biophase-selective histamine H-4 receptor inhibitors are RS-, R- or S-norketotifen, or pharmaceutically acceptable salts thereof, specifically RS-norketotifen or pharmaceutically acceptable salts thereof. In one aspect, the histamine H4-receptor-related disorder is not histamine H-1-related.
Exemplary histamine H-4 receptor-related dermal disorders include histamine H-4 receptor-related pruritic disorders. Activation of dermal histamine H-4 receptors by histamine or other endogenous H-4 receptor agonists, such as for example CCL16, results in pruritus that is caused by pruritic mechanisms that are regulated by histamine H-4 receptors.
In one aspect, a method of treating a mammal in need of treatment for histamine H-4 receptor-related pruritus comprises orally administering to the mammal in need thereof a therapeutically effective amount of a biophase-selective histamine H-4 receptor inhibitor, wherein the biophase-selective histamine H-4 receptor inhibitor is norketotifen, an isomer of norketotifen, or a pharmaceutically acceptable salt thereof. In one embodiment, the histamine H-4 receptor-related pruritus is resistant to treatment with histamine H-1 receptor inhibitors.
Pruritus (itching) is an unpleasant sensation that—contrary to pain—elicits the desire to scratch. Histamine H-4 receptor-related pruritus can be associated with various disorders and conditions, such as for example:
Adverse Effects of Prescription Drugs: antibiotics, antifungals, pain relievers;
Dermal Disorders: autoimmune dermatitis, contact dermatitis, dermal scleroderma, folliculitis, idiopathic dermatitis, infections, insect bites, melanoma, parasites, scabies, sunburn, warts, xerosis and dermal conditions such as moles, hyperpigmentation, hypopigmentation and rashes;
Systemic Disorders: anemia, cholestasis, diabetes, Hodgkin lymphoma, iron deficiency, chronic renal failure, systemic scleroderma, multiple sclerosis, uremia, and conditions such as pregnancy;
Nerve disorders: multiple sclerosis, neuropathic pruritus, scars, shingles;
Psychological disorders: anxiety, depression, emotional stress, neurodermatitis, psychological trauma, psychoses; and
Idiopathic pruritus: various forms of pruritus, where the cause for the itching has not been successfully determined.
Scratching behavior is a sign of pruritus and can be studied in humans and in mice after intradermal injections of histamine. Pruritus following histamine injections is not inhibited by histamine H-1 inhibitors, such as desloratadine (Clarinex® Merck Sharp & Dohme), diphenhydramine (Benadryl® McNeil) or fexofenadine (Allegra® Sanofi Aventis). Histamine-induced pruritus is potently inhibited by histamine H-4 inhibitors, such as for example JNJ7777120 and norketotifen (Example 5).
The methods for treatment of pruritus, disclosed herein, are useful in the treatment of non-humans as well as humans. While it is well known that the human histamine H-4 receptor expresses constitutive activity, it is possible that non-human histamine H-4-receptors do not express constitutive activity. If it is confirmed that the canine and feline histamine H-4 receptor does not express constitutive activity and/or norketotifen therefore proves not to be an inverse histamine H-4 receptor agonist on the canine histamine H-4 receptor, the correct terminology herein shall be “antagonist” or “inhibitor” rather than “inverse agonist”. Therefore, and to improve clarity with regard to histamine H-4 receptors, the terms antagonist, inhibitor and inverse agonist will herein be considered as homologs and will be alternatingly used. The human histamine H-4 receptor has a high homology with the canine histamine H-4 receptor, the homology being 71 percent.
Another advantage of the use of norketotifen and its isomers is that those compounds express inhibitory activities at both histamine H-1 receptors and histamine H-4 receptors. There is a benefit to inhibiting the activities of both H-1 and H-4 receptors, since co-administration of histamine H-1 and histamine H-4 receptor inhibitors can completely inhibit some pruritic effects, such as for example types of pruritus with high intradermal concentrations of histamine. It is believed that the histamine H-4-receptor mediated pruritic activity is initiated locally in the skin, where pruritogenic mediators activate receptors on neuronal C-fibers. However, the mechanisms for the potentiated antipruritic activities of histamine H-1 receptor inhibitors and histamine H-4 receptor inhibitors are presently not understood.
Despite the advantages of combinations of compounds that inhibit histamine H-1 receptors with compounds that inhibit H-4 receptors, no compound with a combination of histamine H-1 receptor and histamine H-4 receptor inhibitory activities has, to our knowledge, been described previously. Norketotifen and the isomers thereof have such combined activities.
Not all types of pruritus are inhibited by histamine H-4 receptor inhibition. Thus, for example, the selective histamine-4 receptor inhibitor JNJ7777120 failed to inhibit pruritus induced by interleukin-31 (IL-31) in a recent study.
Norketotifen can be made from methods known in the art, as described in U.S. Pat. No. 3,682,930, the disclosure of which is hereby incorporated by reference for its teaching of the synthesis of norketotifen.
The norketotifen isomers can be made as described in U.S. Pat. No. 7,226,934 and U.S. Pat. No. 7,557,128, the disclosures of which are hereby incorporated by reference for their teaching of the synthesis of norketotifen isomers.
Norketotifen is the active metabolite of ketotifen, which is a Generation-1 antihistamine. Ketotifen may be the most potent antihistamine ever marketed, but it is also the most sedating of all marketed antihistamines. The sedative effects of ketotifen are strictly dose-limiting and doses higher than 1 mg, bid are rarely used. It is currently believed that approximately 0.5 mg norketotifen is formed in the human body for every 1 mg of ketotifen that is administered. The metabolism—a demethylation of the piperidine nitrogen—takes place in the liver. The sedation by ketotifen is caused by ketotifen per se (the “prodrug”) while the disease-modifying effects of the drug are believed to be due to the metabolite norketotifen. The currently used therapeutic doses of ketotifen—1 mg bid to humans—are not high enough to offer therapeutic activity by inhibition of histamine H-4 receptor activity. (See, e.g., Example 3, Table 3).
Repeat-dose pharmacological and toxicological studies have now been performed in dogs and it has surprisingly been found that daily doses up to 20 mg/kg/day of racemic or isomeric norketotifen can be given chronically to dogs without causing sedation or other adverse events. Doses up to 28 mg/kg/day did not cause sedation and were tolerated by the dogs, but single oral doses of norketotifen of 4 mg/kg to 20 mg/kg are preferred to obtain inhibition of H-4 receptors in the lungs and in the skin of dogs. Dogs vomit spontaneously for a variety of benign reasons—to expel unwanted content from their stomach, as a result of gastric irritation or in response to colonic irritation. In recent toxicological studies in dogs of various drugs, including isomeric norketotifen, vomiting occurred in both control groups and drug groups and may have been treatment-related, rather than drug-related. The unexpectedly low systemic toxicity of norketotifen and the isomers thereof is believed to be due to low systemic (plasma) concentrations, caused by the fact that norketotifen and the isomers thereof are not circulating in plasma but are accumulating in the skin, which is the largest organ in the body.
Similarly, it has been found that doses of 5 mg and 10 mg of norketotifen in humans also do not produce sedation when tested in human subjects.
It is presently estimated that inhibition of histamine H-4 receptor activity at the biophases in mammals suffering from histamine H-4 receptor-related dermal or pulmonary disorders, will occur at oral doses of norketotifen or an isomer thereof from approximately 2 mg/day to approximately 500 mg/day, which leaves a wide safety margin. Useful doses of racemic or isomeric norketotifen to human patients suffering from said disorders are between 2 mg/day and 500 mg/day. More preferred is a daily oral dose of 2 mg/day to 40 mg/day to a human patient and most preferred is a human dose of 2 mg/day to 20 mg/day of norketotifen or an isomers thereof to human patients suffering from histamine H-4 receptor-related dermal or pulmonary diseases.
Useful oral doses of racemic norketotifen or an isomer thereof are 2 mg/kg/day to 28 mg/kg/day to dogs suffering from pruritus diseases. More preferred are oral doses ranging from 4 mg/kg/day to 20 mg/kg/day of racemic norketotifen, or an isomer thereof, to dogs suffering from pruritic diseases.
The embodiments disclosed herein also provide pharmaceutical compositions, which comprise the compound norketotifen, its isomers, and salts, formulated together with one or more pharmaceutically acceptable carriers.
Pharmaceutical compositions for oral administration of solid dosage forms include capsules, granules, pills, powders and tablets. In solid dosage forms, the active compound may be mixed with one or more pharmaceutically acceptable excipients or carriers (such as for example sodium citrate, dicalcium phosphate), fillers or extenders (such as for example starch, lactose, sucrose, glucose, mannitol, silicic acid), binders (such as for example carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, acacia), humectants (such as for example glycerol), solution retarding agents (such as for example paraffin), disintegrating agents (such as for example agar-agar, calcium carbonate, starch, alginic acid, silicates, sodium carbonate), absorption accelerators (such as for example quaternary ammonium compounds), wetting agents (such as for example cetyl alcohol, glycerol monostearate), absorbents (such as for example kaolin, bentonite clay), lubricating agents (such as for example talc, calcium stearate, magnesium stearate, polyethylene glycols, sodium lauryl sulfate), and/or other excipients, such as for example buffering agents.
Solid forms of capsules, granules, pills, and tablets can have coatings and/or shells (such as for example enteric coatings) known in the art. The compositions may also be designed to release the active ingredient(s) in a certain part of the gastrointestinal tract or in a controlled release, slow-release or in a delayed-release manner. The active compound(s) can also be microencapsulated with one or more of the above-mentioned excipients or other suitable excipients.
Liquid dosage forms for oral administration may be preferred administration forms to children suffering from pruritus. Such formulations include for example pharmaceutically acceptable solutions, emulsions, solutions, suspensions, syrups and elixirs. The liquid dosage form may also contain excipients known to those skilled in the art of drug formulations, such as for example diluents (such as for example water, other solvents and solubilizing agents, and mixtures thereof), and emulsifiers (such as for example ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, butylene glycol, dimethyl formamide, oils, oleic acid, glycerol, polyethylene glycols, sorbitan fatty esters, and mixtures thereof. The oral compositions may also include other excipients as known to those skilled in the art.
The compositions described here can also include other drugs with antipruritic activity, as for example a corticosteroid. Due to the antipruritic activity of norketotifen or the isomers thereof, a beneficial steroid-sparing effect will be possible when treating patients suffering from various types of pruritic disorders.
The invention is further illustrated by the following non-limiting examples.
Histamine is excessively released from granulocytes in patients with pruritic diseases. The inhibition of histamine release from human granulocytes (leukocytes; buffy coat) by test articles was studied. Leukocytes were obtained from blood from healthy human volunteers and release of histamine was induced by incubation (20 min/37° C.) of the buffy coats with the calcium ionophore A23187 (5 μM) in the presence or absence of a test article. Histamine was analyzed by enzyme-immune assays, using commercially available kits and a microplate reader (MRX, Dynatech). The test articles were evaluated, in duplicate, at five concentrations.
Norketotifen was approximately 10 times more potent than ketotifen as an inhibitor of histamine release from human inflammatory cells. Thus, norketotifen will potently decrease the concentration of histamine in inflamed tissues.
Affinities of the test compounds for peripheral human histamine H-1-receptors were assessed using receptor-binding assays. The specific binding of the radioactive ligand to the receptor was defined as the difference between total binding and nonspecific binding, determined in the presence of excess unlabeled ligand. Ki-values were determined according to the Cheng-Prusoff equation.
Norketotifen and the isomers thereof had high affinity for histamine H-1-receptors, similar to desloratadine and better than diphenhydramine.
The effects of the reference compounds (ketotifen, loratadine, desloratadine and diphenhydramine) verify the previously known receptor binding activities of these compounds, thereby validating the test methodology.
Affinities of the test compounds for peripheral human histamine H-4-receptors were assessed using receptor-binding assays. The specific binding of the radioactive ligand to the receptor was defined as the difference between total binding and nonspecific binding, determined in the presence of excess unlabeled ligand. [3H]-histamine was used as the ligand in this study and the affinity values were determined according to the Cheng-Prusoff equation.
RS-, S- and R-norketotifen had affinity for the H-4 receptor, albeit lower affinity than the published corresponding values for super-potent and systemically active selective H-4—receptor active compounds, such as for example JNJ 7777120. It is not believed that RS-, S- and R-norketotifen will express systemic reversal of the activity of H-4 receptors unless the compounds appear in high concentrations at the receptor sites.
As known to those skilled in the art of pharmacology, high agonist concentrations are needed in these studies since the Cheng-Prusoff equation is resulting in erroneous values at low concentrations of agonists.
It is believed to be of therapeutic importance that norketotifen inhibits both histamine H-4 and histamine H-1 receptors since a potentiation of the antipruritic activity of histamine H-4 inhibition by histamine H-1 inhibition has been reported in the art, although histamine H-1 inhibition by itself has no antipruritic effects, as described in the art and as shown by the present results (Example 5;
Five male beagle dogs, weighing 11.2-13.9 kg (2-4 years old) were used in the study. All animals were administered gelatin capsules containing oral doses of the test article 8.0 mg/kg/day as a hydrogen fumarate salt, equal to 5.6 mg/kg/day of the free base. The animals were dosed once daily for four consecutive weeks followed by daily observations for an additional two-week washout period.
Multiple plasma samples and skin biopsies were taken from each dog on Day 1 and Day 28 of drug administration. The plasma and skin samplings were performed at pre-dose, and at 2, 6, 12 and 24 hours post-dose. Plasma and skin samples were also taken intermittently at predetermined intervals during the 28-days dosing period and up to the last day of the study, which was Day 42. Blood samples were taken from v. cephalica antebrachii. Skin biopsies were taken from the area between the mid ventral to lateral abdominal areas, using a 6 mm (diameter) skin biopsy device (Acu-Punch®, Acuderm® Inc., Fort Lauderdale, Fla. 33309). Multiple plasma and biopsy samples were obtained from each of 4 or 5 dogs. Subcutaneous fat deposits were carefully trimmed from the skin samples and the skin samples were weighed. All plasma and skin samples were kept
The plasma samples and skin biopsy samples were analyzed using LC/MS/MS methodology. All pharmacokinetic analyses were performed using Pharsight WinNonlin® Professional v5.2.1 software.
Since norketotifen potently inhibits the release of histamine from pro-inflammatory cells (Example 1) and acts as an inverse agonist at the histamine H-4 receptors (Example 3), and since norketotifen is accumulated in the skin (Table 4), those skilled in the art will realize that the decreased availability of histamine together with the inhibitory activity at the histamine H-4 receptor site will act synergistically to reduce the G-protein-mediated signaling from dermal histamine H-4 receptors.
Antipruritic effects were tested in vivo in CD-1 mice, females, 10-12 weeks, according to methods known in the art. The hair was clipped over the rostral part of the back at the interscapular level of the mice one day before the dosing. Before the testing, the mice were placed in individual clear plastic cages for at least one hour for acclimation. After fasting for 1.5 hours, the animals were dosed orally with the test article, dissolved in a vehicle consisting of 1% methylcellulose/water, 10 mL/kg body weight. Sixty minutes after the oral dosing, an intradermal injection of histamine (300 nmol in 20 μl phosphate buffered saline (PBS), pH 7.4) was administered. Immediately after the histamine injection, the bouts of scratches were counted for 40 min. Scratching induced by the histamine vehicle PBS served as control.
Norketotifen was tested in escalating doses and in a supramaximal dose of 100 mg/kg. JNJ7777120 and desloratadine were dosed orally with 20 mg/kg, which is within the murine dose ranges used for those compounds in the art. The vehicle for the reference compound JNJ7777120 was 20% hydroxypropyl-B-cyclodextrin in water. The vehicle for the reference compound desloratadine was the same as the vehicle for norketotifen (10 mL/kg of 1% methylcellulose in water). The test results are shown in
Norketotifen was dose-dependently inhibiting histamine-induced pruritus and a supramaximal dose (100 mg/kg) demonstrated complete inhibition. Scratches induced by the histamine-free vehicle PBS (not shown in
It was concluded that norketotifen potently and dose-dependently decreased histamine-induced pruritus. The results from tests of a selective histamine-4 inhibitor and a selective histamine-1 inhibitors demonstrate that histamine H-4 inhibition, but not histamine H-1 inhibition blocks histamine—induced pruritus. The unexpectedly potent inhibition of pruritus by norketotifen may in part be due to potentiation caused by simultaneous expression of both histamine H-1 and histamine H-4 receptor inhibition by norketotifen.
Formulations for oral administration of norketotifen (such as for example tablets, capsules and syrups) have been developed.
The active ingredient is blended with the lactose and cellulose until a uniform blend is formed. The blue lake is added and further blended. Finally, the calcium stearate is blended in, and the resulting mixture is compressed into tablets using for example a 9/32-inch (7 mm) shallow concave punch. Tablets of other strengths may be prepared by altering the ratio of active ingredient to the excipients or to the final weight of the tablet.
Those skilled in the art realize that oral formulations can be in the form of, for example, a tablet, a capsule, a dog-treat, a cat-treat, a syrup or another form of liquid formulation.
As used herein, the terms “pharmaceutically acceptable salts” or “a pharmaceutically acceptable salt thereof” refer to norketotifen salts, which have been prepared from pharmaceutically acceptable non-toxic acids. Exemplary pharmaceutically acceptable acid addition salts for the compound of the present invention include acetic, benzenesulfonic (besylate), benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pathothenic, phosphoric, p-toluenesulfonic, succinic, sulfuric, tartaric, and the like. The hydrochloride salt and the hydrogen fumarate salt are particularly preferred.
The use of the terms “a” and “an” and “the” and similar referents (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The terms first, second etc. as used herein are not meant to denote any particular ordering, but simply for convenience to denote a plurality of, for example, layers. The terms “comprising”, “having”, “including”, and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to”) unless otherwise noted. As used herein, the term mammal includes humans, dogs, and cats. Recitation of ranges of values are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. The endpoints of all ranges are included within the range and independently combinable. All methods described herein can be performed in a suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”), is intended merely to better illustrate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention as used herein.
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. In addition, many modifications may be made to adapt a particular situation or material to the teachings of the invention without departing from essential scope thereof. Therefore, it is intended that the invention not be limited to the particular embodiment disclosed as the best mode contemplated for carrying out this invention, but that the invention will include all embodiments falling within the scope of the appended claims.
