HK1110213A - Compositions for treatment of inflammation and pain using a combination of a cox-2 selective inhibitor and a ltb4 receptor antagonist - Google Patents

Description

Use of COX-2 selective inhibitors and LTB4Combination of receptor antagonists for the treatment of inflammation and pain

Background

Technical Field

The invention relates to a method for preparing a compound for relieving inflammation by using non-steroidal anti-inflammatory compounds and leukotriene B₄Combinations of receptor antagonists for use in compositions and methods for treating and preventing pain, pain-related disorders, inflammation, and inflammation-related disorders.

Background

Inflammatory mediators (inflammation mediators) have been implicated as playing a major pathogenic role in causing, spreading and prolonging pain and inflammation. Prostaglandins have been shown to be important mediators of inflammation and modulators of other important functions not directly related to inflammation. Modulation of prostaglandin production and activity has become a common goal in anti-inflammatory drug development activities. However, common non-steroidal anti-inflammatory drugs (NSAIDs) that are active in reducing prostaglandin-induced pain and swelling associated with inflammatory processes sometimes have adverse effects on other prostaglandin-regulated processes that are not associated with inflammatory processes.

The mechanism of action of some common NSAIDs is to regulate prostaglandin synthesis by inhibiting the cyclooxygenase enzyme that catalyzes the conversion of arachidonic acid (i.e., the first step in the prostaglandin synthesis pathway). In the eighties of the twentieth century, Needleman et al discovered two epoxidases associated with the above transformations. These enzymes have been referred to as cyclooxygenase-1 (herein referred to as "COX-1") and cyclooxygenase-2 (herein referred to as "COX-2"). See Needleman, p, et al, j.rheumato, 24, supplement 49: 6-8(1997).

COX-1 has been shown to be an enzyme that grows without physiological requirements or substrate concentrations and is associated with some of the non-inflammatory regulatory functions associated with prostaglandins. On the other hand, COX-2 is an inducible enzyme apparently involved in the inflammatory process. Inflammatory stimuli cause COX-2 to be produced, stimulating the release of prostaglandins. In turn, prostaglandins sensitize peripheral nociceptive terminals, causing hypersensitivity to local pain. Some common NSAIDs are now known to be in vitro inhibitors of both COX-1 and COX-2. Thus, when administered at sufficiently high levels of these NSAIDs, the NSAIDs not only affect the inflammatory consequences of COX-2 activity, but also affect the beneficial activity of COX-1.

Compounds have been found to selectively inhibit COX-2 in an in vitro enzyme assay. Novel COX-2 selective inhibitors offer advantages including the ability to inhibit or reduce inflammation while avoiding the adverse side effects associated with COX-1 inhibition. Thus, COX-2 selective inhibitors have broad application prospects in therapies involving chronic dosing, such as for the management of pain and inflammation in arthritis.

Leukotriene (Leukotriene) is a compound produced by the metabolism of arachidonic acid in mammals. Arachidonic acid is metabolized in mammals through two distinct pathways, one producing prostaglandins and thromboxanes, and the other producing oxidation products known as leukotrienes. There are several different classes of leukotrienes, including leukotriene A₄Leukotriene B₄(referred to herein as LTB)₄) Leukotriene C₄And leukotriene D₄. It is believed that LTB₄Is a mediator of inflammation and plays a major role in, for example, arthritis, psoriasis, myocardial infarction, irritable bowel disease and other diseases.

In contrast to other leukotrienes, which primarily cause stimulatory contraction and some proinflammatory effects, LTB₄Mainly as chemotactic and activating agents for leukocytes (Jennewein, H.M., et al, Prog respir. Res. Basel. Karger, 2001, vol 31, pp 121-125).

LTB₄Receptors are located on a variety of cells, primarily neutrophils, and also macrophages, lymphocytes, eosinophils, and lung epithelial cells. In the case of segmented-leaf-nucleus leukocytes (PMNLs), LTB₄Causing chemotaxis, chemical movement, oxidative burst and up-regulation or shedding of adhesion molecules as a necessary condition for adhesion. The LTB₄Neutrophil apoptosis is also inhibited, thereby prolonging the inflammatory response.

LTB₄Plays an important role in the pathophysiology of rheumatoid arthritis (herein referred to as "RA") (Alten R. et al in Ann Rheum Dis.2004, Feb.; 63 (2): 170-6). RA is an autoimmune disease characterized by joint inflammation, joint destruction, antegrade disability, and premature death. The dire consequences of RA can be minimized by treating RA patients with disease-modifying antirheumatic drugs, either alone or in combination with other drugs. Treatment with disease-modifying antirheumatic drugs to limit or inhibit other diseasesDevelopment (Hochberg, M.C., Scan JRheumatotol subcategory 1999; 112: 3-7)

US patent 5384318 describes certain substituted sulphonamide drugs which are LTB₄An antagonist.

U.S. Pat. No. 5246965 describes aromatic ethers which are LTB₄A receptor antagonist.

US patent 6342510 describes COX-2 inhibitors and LTB₄Combinations of receptor antagonists are useful in treating inflammation and disorders associated with inflammation.

PCT patent application WO 2004/047824 describes certain pharmaceutical compositions comprising LTB having a hydroxyl group and a benzamidine group₄A receptor antagonist; and certain COX-2 inhibitors; and a pharmaceutically acceptable excipient.

There is a need for a therapy that inhibits pain and inflammation and also has a disease modifying effect.

Disclosure of Invention

It may be noted that several embodiments of the present invention provide a therapeutic composition comprising at least one COX-2 selective inhibitor or prodrug thereof (produgs) in combination with at least one LTB₄A receptor antagonist, wherein the LTB₄The receptor antagonist comprises one or more compounds selected from the group consisting of 2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl]-4- (trifluoromethyl) benzoic acid, pharmaceutically acceptable salts thereof and mixtures thereof.

In another embodiment, the present invention provides a method for treating, preventing or inhibiting inflammation, an inflammation-related disorder, pain or a pain-related disorder in a subject in need thereof, said method comprising administering to said subject a composition comprising at least one COX-2 selective inhibitor or prodrug thereof and at least one LTB₄Treatment with receptor antagonistsComposition wherein the LTB₄The receptor antagonist comprises one or more compounds selected from the group consisting of 2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl]-4- (trifluoromethyl) benzoic acid, pharmaceutically acceptable salts thereof and mixtures thereof.

Further areas of applicability of the present invention will become apparent from the detailed description provided hereinafter. It should be understood, however, that the detailed description and the examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

Drawings

FIG. 1 murine Air Pouch 2h zymosan

FIG. 2 disease severity

FIG. 3 incidence of collagen-induced arthritis

FIG. 4 combination reduces the incidence of collagen-induced arthritis

Detailed Description

The detailed description provided below helps those skilled in the art practice the present invention. Nevertheless, these detailed descriptions should not be construed as unduly limiting the invention since modifications and variations in the embodiments discussed herein may be made by persons of ordinary skill in the art without departing from the spirit or scope of the present disclosure.

The contents of each of the references cited herein, including the contents of the references cited in these primary references, are hereby incorporated by reference in their entirety.

a. Definition of

The following definitions are provided to assist the reader in understanding the detailed description of the invention:

as used herein, the term "COX-1" refers to one of the two isomeric reformulations of the fatty acid epoxidase known as cyclooxygenase-1.

The term "COX-2" as used herein refers to one of two isomeric reformates of a fatty acid epoxidase known as cyclooxygenase-2.

The term "mg" as used herein refers to milligrams.

The term "g" as used herein refers to grams.

The term "mpk" as used herein means milligrams per kilogram

The term "SLS" as used herein means sodium lauryl sulfate.

The term "PVP" as used herein means polyvinylpyrrolidone.

The term "inflammation-associated disorder" or "inflammatory disorder" as used herein is meant to include, but is not limited to, the individual symptoms or disorders referred to below. However, these terms are also meant to include any therapeutic situation in which inflammation or processes associated with inflammation play a role.

The terms "neoplasia" and "neoplasia disorder" (used interchangeably herein) refer to any new and abnormal cell growth, including any in which cell proliferation is uncontrolled and more severe. Tumors can be benign or malignant. Neoplasia also includes the term "cancer," and for purposes of the present invention, cancer is a sub-type of neoplasia. The term "neoplastic disorder" as used herein also includes other cellular abnormalities, such as, for example, hyperplasia, metaplasia and dysplasia. The terms neoplasia, metaplasia, dysplasia and hyperplasia may be used interchangeably herein and generally refer to cells that undergo abnormal cell growth.

The two terms "neoplasia" and "neoplasia disorder" refer to "tumors" or neoplasms, which may be benign, precancerous, metastatic, or malignant.

The term "prevention" as used herein refers to any means (whether slight) to improve a subject's predisposition to a disease or disorder or to reduce the risk of developing a disease or disorder, including any of the following: (1) substantially preventing the onset of a clinically significant disorder or disease in a subject of interest; (2) preventing the onset of a disease or disorder in a target subject at a preclinically significant stage; or (3) substantially preventing the disorder or disease in the subject. This definition includes prophylactic treatment.

The term "inhibit" as used herein means to reduce the severity of a disorder or disease as compared to the disorder or disease that occurs without the application of the present invention. This reduction in severity can result from a reduction in any one or more of the symptomatic characteristics of the disease or disorder. For example, in the case of inflammation, the symptom characteristics include, for example, swelling pain, redness, stiffening, and others. At the cellular level, it comprises features such as: chemotaxis; inflammatory mediators such as prostaglandins, chemokines, leukotrienes are released; cell infiltration; activate immune cells and others.

The phrase "therapeutically effective" means that the amount of each agent is limited such that the method will achieve the goal of improving the severity and frequency of the disorder relative to untreated or treatment with each agent alone, while avoiding the adverse side effects normally associated with replacement therapy.

For therapeutic or prophylactic purposes, the term "subject" includes any human or animal subject susceptible to a disorder or disease. The target subject may be a domestic animal, a laboratory animal, a zoo animal, or a companion animal. In one embodiment, the target subject is a mammal. In an alternative embodiment, the mammal is a human. In another embodiment, the target subject is a companion animal such as a dog, cat, or horse.

The term "COX-2 selective inhibitor" includes compounds that selectively inhibit the COX-2 enzyme over the COX-1 enzyme, and also includes pharmaceutically acceptable salts and prodrugs of such compounds.

The selectivity of a COX-2 inhibitor varies depending on the conditions under which the test is performed and the inhibitor being tested. For illustrative purposes, the selectivity of COX-2 inhibitors may be based on the IC of COX-1 inhibition in vitro or in vivo₅₀IC of value and COX-2 inhibitory effect₅₀Ratio of values (COX-1 IC)₅₀/COX-2 IC₅₀) To measure. The COX-2 selective inhibitor is any COX-1IC₅₀With COX-2 IC₅₀Inhibitors with a value greater than 1. In one embodiment of the invention, the ratio is greater than 2. In another embodiment of the present invention, the ratio is greater than 5. In another embodiment of the present invention, the ratio is greater than 10. In another embodiment, the ratio is greater than 50. In another embodiment of the present invention, the ratio is greater than 100. For details on the determination of COX-2 inhibitor activity and COX-2 selectivity in vitro, see example 1.

For COX-2 selective inhibitors, the term "IC₅₀"refers to the concentration of compound required to produce 50% inhibition of enzyme activity in an in vitro enzyme assay as described below. In one embodiment of the invention, the COX-2 selective inhibitor has an IC of less than about 1 micromolar₅₀Alternatively less than about 0.5 micromolar and alternatively less than about 0.2 micromolar.

For LTB₄Receptor antagonists, the term "IC₅₀"means a specificity sufficient to inhibit 50% in an in vitro assay as described below³H-LTB₄The concentration of the bound compound. In one embodiment of the invention, LTB₄The receptor antagonist has an IC of less than about 1 micromolar₅₀Alternatively less than about 0.5 micromolar and alternatively less than about 0.2 micromolar.

In one embodiment of the invention, the COX-2 selective inhibitor has an IC of greater than about 1 micromolar₅₀. In another embodiment, the COX-2 selective inhibitor has an IC of greater than 20 micromolar₅₀. Inhibitors of the cyclooxygenase pathway in the metabolism of arachidonic acid for use in the present methods of the invention may exhibit enzymatic activity by a variety of mechanisms. By way of example and not limitation, asThe inhibitors in the methods described herein may directly block the activity of the enzyme by acting as an enzyme medium.

The term "leukotriene B" as used herein₄Receptor antagonist "or" LTB₄Receptor antagonist "or" LTB₄ra' includes selective inhibition of LTB₄Having an IC of less than about 10 micromolar for the acceptor₅₀The compound of (1). In another embodiment of the invention, LTB₄The receptor antagonist has an IC of less than about 1 micromolar₅₀. Example 2 illustrates the use in this application for determining LTB₄Methods of receptor antagonistic activity.

The term "treatment" as used herein means alleviation of symptoms, temporary or permanent elimination of the causative agent or alteration or slowing of the appearance or worsening of symptoms. The term "treating" includes, but is not limited to, ameliorating or inhibiting the pathogenesis of any of the disorders, diseases, symptoms associated with the disorder or disease-related condition described herein.

Also included within the scope of the present invention are prodrugs or LTB which are COX-2 selective inhibitors₄A compound that is a prodrug of a receptor antagonist. The term "prodrug" as used herein with reference to a COX-2 selective inhibitor refers to a compound that can be converted in the body of a subject of interest to an active COX-2 selective inhibitor by a metabolic process or by a simple chemical process. An example of a prodrug of a COX-2 selective inhibitor is celecoxib (parecoxib), which is a therapeutically effective prodrug of the COX-2 selective inhibitor valdecoxib (valdecoxib). An example of a preferred COX-2 selective inhibitor prodrug is tebufenpyrad. A class of prodrugs of COX-2 inhibitors is described in US patent 5932598.

The term "pharmaceutically acceptable" as used herein means that the noun modified by it applies to pharmaceutical products.

As used herein, "effective amount" means the dose or amount administered to a target subject and the frequency of administration to a target subject, which is readily determined by one of ordinary skill in the art using known techniques and observing results obtained in similar circumstances.

The term "subject in need of such treatment or prevention" as used herein means any subject that has the disease or disorder being treated during survival or is at risk of developing the disease or disorder or symptoms of the disease or disorder.

b. Details of

In accordance with the present invention, there is now disclosed a therapeutic composition comprising a COX-2 selective inhibitor or prodrug thereof and an LTB₄A receptor antagonist, wherein the LTB₄The receptor antagonist is selected from the group consisting of:

2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl ] -4- (trifluoromethyl) benzoic acid;

4- [ [3- [ [4- [1- (4-hydroxyphenyl) -1-methylethyl ] phenoxy ] methyl ] phenyl ] methoxy ] benzamidine;

[ [4- [ [3- [ [4- [1- (4-hydroxyphenyl) -1-methyl-ethyl ] phenoxy ] methyl ] -phenyl ] methoxy ] -phenyl ] iminomethyl ] carbamic acid ethyl ester;

4- [1- [4- [ [3- [ [4- (aminoiminomethyl) phenoxy ] -methyl ] phenyl ] methoxy ] phenyl ] -1-methylethyl ] phenyl β -D-glucopyranosiduronic acid;

2- [ (3R, 4S) -3- (1, 1' -biphenyl-4-ylmethyl) -4-hydroxy-3, 4-dihydro-2H-chromen-7-yl ] -4- (trifluoromethyl) benzoic acid;

2- ((3S) -3-benzyl-3, 4-dihydro-4-hydroxy-2H-chromen-7-yl) -4-chlorobenzoic acid;

2- ((3R) -3-benzyl-3, 4-dihydro-3, 4-dihydroxy-2H-chromen-7-yl) -4-fluorobenzoic acid;

1-carboxy-1- (((3S, 4S) -3, 4-dihydroxy-3- (((4- (phenyl) -methyl) -chromen-7-yl)) cyclopentane, the pharmaceutically acceptable salts thereof, and mixtures of any of these compounds or salts.

Some LTBs that can be used in the present invention₄The receptor antagonists are listed in table 1. Pharmaceutically acceptable salts of the compounds listed in table 1 may also be used in the present invention.

TABLE 1 LTB₄Receptor antagonists

In one embodiment of the invention, LTB₄The receptor antagonist comprises 2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl]-4- (trifluoromethyl) benzoic acid or a salt thereof.

Some of the compounds that may be used as COX-2 selective inhibitors in the present invention are individually listed in Table 2. These compounds may be used in the present invention alone or in combination with two or more COX-2 selective inhibitors. In addition, pharmaceutically acceptable salts and prodrugs of the compounds listed in table 2 may also be used in the present invention.

TABLE 2 some useful COX-2 inhibitors

In one embodiment of the invention, a therapeutic composition comprises any LTB of table 1₄A receptor antagonist, a prodrug, or a salt thereof and any of the COX-2 inhibitors, prodrugs, or salts thereof in table 2.

In one embodiment, the COX-2 selective inhibitor includes one or more diaryl heterocyclic COX-2 selective inhibitors.

Alternatively, the COX-2 selective inhibitor may be a benzopyran COX-2 selective inhibitor. US patent 6024356 (incorporated herein by reference) describes some useful selective inhibitors of benzopyran COX-2. Other useful COX-2 selective inhibitors are described in US patent application 10/801446 (incorporated herein by reference). Other useful COX-2 selective inhibitors are described in US patent application 10/801429 (incorporated herein by reference).

In one embodiment, the COX-2 inhibitor comprises celecoxib.

In another embodiment of the invention, the COX-2 selective inhibitor is celecoxib, LTB₄The receptor antagonist is 2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl]-4- (trifluoromethyl) benzoic acid (compound L1). ReplaceableA pharmaceutically acceptable salt of compound L1 may be used in the present invention. For example, a useful pharmaceutically acceptable salt of compound L1 is the ethylenediamine salt of L1, e.g., the mono (ethylenediamine) salt of L1. Salts and polymorphs of L1 useful in the present invention are described in US patent 6436987, which is incorporated herein by reference. In another embodiment of the invention, the COX-2 selective inhibitor is valdecoxib, LTB₄The receptor antagonist is compound L1.

Another embodiment of the present invention provides a method for treating, preventing or inhibiting inflammation, an inflammation-related disorder, a pain-related disorder or pain in a subject in need thereof, comprising administering to the subject a composition comprising at least one COX-2 selective anti-inflammatory compound, salt, prodrug or mixture thereof and at least one LTB₄A receptor antagonist, a salt, a prodrug, or a mixture thereof.

Another embodiment of the present invention provides a method for treating, preventing or inhibiting inflammation, an inflammation-related disorder, a pain-related disorder or pain in a subject in need thereof, comprising administering to the subject a composition comprising at least one COX-2 selective inhibitor (or salt, prodrug or mixture thereof) selected from Table 2 and at least one LTB selected from Table 1₄A receptor antagonist (or a salt or a mixture thereof). For example, the present invention provides a method for treating, preventing or inhibiting inflammation. In another embodiment, the invention provides a method for treating, preventing or inhibiting a disorder associated with inflammation.

The present invention also provides a method for treating, preventing or inhibiting inflammation, an inflammation-related disorder, a pain-related disorder or pain in a subject in need thereof, wherein the method comprises administering to the subject a composition comprising at least one COX-2 selective anti-inflammatory compound, salt, prodrug, or mixture thereofSubstance and at least one LTB₄A receptor antagonist compound, wherein the LTB₄The receptor antagonist comprises one or more compounds selected from the group consisting of 2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl]-4- (trifluoromethyl) benzoic acid, salts thereof and mixtures thereof.

Another embodiment of the present invention is a method for treating, preventing or inhibiting inflammation, an inflammation-related disorder, a pain-related disorder or pain in a subject in need thereof, comprising administering to the subject a composition comprising celecoxib or a salt or prodrug or a mixture thereof and an LTB₄A receptor antagonist compound.

Another embodiment of the present invention is a method for treating, preventing or inhibiting inflammation, an inflammation-related disorder, a pain-related disorder or pain in a subject in need of such treatment, prevention or inhibition, the method comprising administering to the subject a composition comprising celecoxib, or a salt, a prodrug, or a mixture thereof, and 2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl ] -4- (trifluoromethyl) benzoic acid, or a prodrug or a salt and a mixture thereof.

In another embodiment, the present invention provides a therapeutic composition comprising a COX-2 selective inhibitor or prodrug thereof and LTB₄A receptor antagonist, wherein the LTB₄The receptor antagonist comprises 2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl]-4- (trifluoromethyl) benzoic acid (compound L1) or one or more salts thereof or mixtures thereof. Some useful forms of the L-1 compounds are described in US patent 6436987, which is incorporated herein by reference. Each of the compounds of table 1 may be used in its acid or base (e.g., conjugate acid or conjugate base) form or in any of its pharmaceutically acceptable salt forms. For example, compound L-1 can be used in any of its pharmaceutically acceptable salt forms. These salts include ethylenediamine salts and crystals of ethylenediamine saltsAnd (3) a body. In addition, each compound of table 1 may be used in any of its crystalline forms (or mixtures thereof) or in an amorphous form. For example, some useful crystalline forms of compound L-1 are described in US patent 6435987.

The composition of the present invention may utilize the LTB described above₄Any one or more of the receptor antagonists in combination with any one or more COX-2 selective inhibitors. For example, the compositions of the present invention may utilize the LTB described above₄Any one or more of the receptor antagonists in combination with any one or more of the COX-2 selective inhibitors described above. Table 3 lists LTB which can be used in the present invention₄Some combinations of receptor antagonists with COX-2 selective inhibitors.

TABLE 3

Example No. 2	COX-2 selective inhibitors	LTB4Receptor antagonists
			1	Celecoxib	L-1
2	Celecoxib	L-2
			3	Celecoxib	L-3
4	Celecoxib	L-4
			5	Celecoxib	L-5
6	Celecoxib	L-6
			7	Celecoxib	L-7

8	Celecoxib	L-8
			9	Celecoxib	L-9
10	Valdecoxib	L-1
			11	Valdecoxib	L-2
12	Valdecoxib	L-3
			13	Valdecoxib	L-4
14	Valdecoxib	L-5
			15	Valdecoxib	L-6
16	Valdecoxib	L-7
			17	Valdecoxib	L-8
18	Valdecoxib	L-9
			19	Afraid of remoxib	L-1
20	Afraid of remoxib	L-2
			21	Afraid of remoxib	L-3
22	Afraid of remoxib	L-4
			23	Afraid of remoxib	L-5
24	Afraid of shakeCloth	L-6
			25	Afraid of remoxib	L-7
26	Afraid of remoxib	L-8
			27	Afraid of remoxib	L-9
28	Delaxib	L-1
			29	Delaxib	L-2

30	Delaxib	L-3
			31	Delaxib	L-4
32	Delaxib	L-5
			33	Delaxib	L-6
34	Delaxib	L-7
			35	Delaxib	L-8
36	Delaxib	L-9
			37	Rofecoxib	L-1
38	Rofecoxib	L-2
			39	Rofecoxib	L-3
40	Rofecoxib	L-4
			41	Rofecoxib	L-5
42	Rofecoxib	L-6
			43	Rofecoxib	L-7
44	Rofecoxib	L-8
			45	Rofecoxib	L-9
46	Etoxicib	L-1
			47	Etoxicib	L-2
48	Etoxicib	L-3
			49	Etoxicib	L-4
50	Etoxicib	L-5
			51	Etoxicib	L-6

52	Etoxicib	L-7
			53	Etoxicib	L-8
54	Etoxicib	L-9
			55	Romexib	L-1
56	Romexib	L-2
			57	Romexib	L-3
58	Romexib	L-4
			59	Romexib	L-5
60	Romexib	L-6
			61	Romexib	L7
62	Romexib	L-8
			63	Romexib	L--9
64	Compound C-8	L-1
			65	Compound C-8	L2
66	Compound C-8	L-3
			67	Compound C-8	L-4
68	Compound C-8	L-5
			69	Compound C-8	L6
70	Compound C-8	L-7
			71	Compound C-8	L-8
72	Compound C-8	L9
			73	Compound C-9	L-1

74	Compound C-9	L-2
			75	Compound C-9	L-3
76	Compound C-9	L-4
			77	Compound C-9	L-5
78	Compound C-9	L-6
			79	Compound C-9	L-7
80	Compound C-9	L-8
			81	Compound C-9	L-9
82	Meloxicam	L-1
			83	Meloxicam	L-2
84	Meloxicam	L-3
			85	Meloxicam	L-4
86	Meloxicam	L-5
			87	Meloxicam	L-6
88	Meloxicam	L-7
			89	Meloxicam	L-8
90	Meloxicam	L-9
			91	Cimicoxib	L-1
92	Cimicoxib	L-2
			93	Cimicoxib	L-3
94	Cimicoxib	L-4
			95	Cimicoxib	L-5
96	Cimicoxib	L-6

97	Cimicoxib	L-7
			98	Cimicoxib	L-8
99	Cimicoxib	L-9

Pharmaceutically acceptable salts may be used in the compositions of the invention for a variety of reasons, including the aqueous solubility of the pharmaceutically acceptable salt. These salts must have a pharmaceutically acceptable anion or cation. Exemplary pharmaceutically acceptable salts can be prepared from the following acids: formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, gluconic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, glucuronic acid, maleic acid, fumaric acid, pyruvic acid, aspartic acid, glutamic acid, benzoic acid, aminobenzoic acid, methanesulfonic acid, stearic acid, salicylic acid, p-hydroxybenzoic acid, phenylacetic acid, mandelic acid, Embonic acid (pamoic acid), methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, pantothenic acid, toluenesulfonic acid, 2-hydroxyethanesulfonic acid, sulfanilic acid, cyclohexylsulfamic acid, alginic acid, b-hydroxybutyric acid, mucic acid, and galacturonic acid.

Suitable pharmaceutically acceptable base addition salts of the compounds of the present invention include metal ion salts and organic ion salts. In some embodiments, metal ion salts include, but are not limited to, the appropriate alkali metal (group Ia) salts, alkaline earth metal (group IIa) salts, and other physiologically acceptable metal ions. Such salts may be prepared from ions of aluminum, calcium, lithium, magnesium, potassium, sodium and zinc. In some cases, organic salts can be prepared from primary, secondary, tertiary amines, and quaternary ammonium salts, the base moieties including trimethylamine, diethylamine, N' -dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine), and procaine. All the salts mentioned above can be prepared in a conventional manner by the person skilled in the art from the corresponding compounds of the invention.

Pharmaceutically acceptable cations include metal ions and organic ions. For example, useful metal ions include, but are not limited to, suitable alkali metal salts, alkaline earth metal salts, and other physiologically acceptable metal ions. Exemplary ions include aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc of conventional valency. In some embodiments, the organic ions comprise protonated primary, secondary, tertiary amines, and quaternary ammonium cations, including, in part, trimethylamine, diethylamine, N' -dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine), and procaine.

Exemplary pharmaceutically acceptable acids include, but are not limited to, hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, methanesulfonic acid, acetic acid, formic acid, tartaric acid, maleic acid, malic acid, citric acid, isocitric acid, succinic acid, lactic acid, gluconic acid, glucuronic acid, pyruvic acid, oxalic acid, oxaloacetic acid, fumaric acid, propionic acid, aspartic acid, glutamic acid, benzoic acid, and the like.

The pharmaceutical compositions of the present invention may be administered in a variety of forms. For example, the composition may be in the form of: tablets, lozenges, sachets, capsules, chewing gum, chewable tablets, controlled release formulations, sustained release formulations, fast dissolving films, gels (e.g., gel capsules), semisolids, solutions (aqueous or non-aqueous), suspensions, intimate mixtures of components (an immediate mix), or any combination of two or more of the foregoing.

In one embodiment of the invention, the COX-2 inhibitor and LTB₄The inhibitor composition is in solid dosage form. For example, the solid dosage form may be an oral dosage form. In another embodiment, the oral dosage form is selected from the group consisting of tablets, capsules, suppositories, pills, gel caps and granules. In another embodiment, the oral dosage form is a capsule. In another embodiment, the capsule is a time release capsule. Such controlled release capsules may, for example, release the active ingredient from the matrix, and in another example, they may release the active ingredient from a controlled release matrix mixture at different rates. In another embodiment, the oral dosage form is a tablet dosage form. In another embodiment, the tablet dosage form may be, for example, a multilayer tablet dosage form (e.g., a single layer for each active ingredient), a disk, a sustained release tablet dosage form, a core shell tablet dosage form, and a side-by-side tablet dosage form (e.g., one side for each active ingredient). In another embodiment, the tablet dosage form comprises a multilayer tablet dosage form. In another embodiment, the tablet dosage form comprises a side-by-side tablet dosage form. In another embodiment, the tablet dosage form comprises a sustained release tablet dosage form. In another embodiment, the tablet dosage form comprises a core shell tablet dosage form. In another embodiment of the invention, the tablet dosage form comprises an osmotic tablet having a core containing one drug and a coating containing another drug. The osmotic tablets may also be contained in a coreAnd another component in the coating, such as an excipient or otherwise.

Pharmaceutical compositions suitable for oral administration may be presented as discrete units, e.g., capsules, sachets, lozenges, or troches, each of which contains a predetermined amount of at least one therapeutic compound useful in the present invention; such as powders or granules; e.g., solutions or suspensions in aqueous or non-aqueous liquids; or for example an oil-in-water emulsion or a water-in-oil emulsion. As indicated, such compositions are prepared by any suitable pharmaceutical method comprising the steps of: the active compound is combined with a carrier, which may be composed of one or more additional ingredients. In general, the compositions are prepared by the following steps: the active compound is mixed homogeneously and intimately with liquid or finely divided solid carriers or both, and the product is then shaped, if necessary. For example, a tablet may be prepared by: the compound powder or granules are optionally compressed or shaped with one or more additional ingredients. Compressed tablets may be prepared by the following steps: the compound in free-flowing form, e.g. as a powder or granules, is compressed in a suitable machine with optional binders, lubricants, inert diluents and/or surfactants/dispersants. Shaped tablets may be prepared by shaping in a suitable machine a powdered compound moistened with an inert liquid diluent.

Comprising a COX-2 selective inhibitor and LTB₄Syrups and elixirs of receptor antagonists may be formulated with sweetening agents, for example glycerol, sorbitol or sucrose. Such formulations may also contain analgesics, preservatives and flavouring and colouring agents. Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs containing inert diluents commonly used in the art, such as water. Such compositions may also include adjuvants, such as wetting agents, emulsifying and suspending agents, and sweetening, flavoring, and perfuming agents.

The invention also encompasses oral administration or "sublingual" buccal administration, and dosage forms include lozenges or chewing gums comprising a compound as set forth herein. The compound may be deposited in a flavouring base, typically sucrose and gum arabic or tragacanth, and pastilles comprising the compound in an inert matrix such as gelatin and glycerol or sucrose and gum arabic.

Pharmaceutical compositions suitable for parenteral administration may generally comprise sterile aqueous preparations of the compounds of the invention. These formulations may be administered intravenously, but administration may also be achieved by subcutaneous, intramuscular or intradermal injection or by infusion. Such formulations are generally prepared by the following steps: the compound is mixed with water and the resulting solution is sterilized and isotonic with blood. The injectable composition according to the invention typically comprises 0.1-10% w/w of the compounds disclosed herein.

Injectable preparations (e.g., sterile injectable aqueous or oleaginous suspensions) may be formulated according to the prior art using suitable dispersing or hardening agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a parenterally-acceptable sterile diluent or solvent, for example, as a solution in 1, 3-butanediol. Acceptable vehicles and solvents that can be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, non-volatile oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

COX-2 inhibitors and LTB₄Administration of one or both of the receptor antagonists may be by inhalation in the form of an aerosol or nebulizer solution. Thus, in one embodiment, a COX-2 inhibitor and LTB₄The receptor antagonist is delivered by direct inhalation, administered into the respiratory system of the target subject as a mist or other aerosol or dry powder.

Pharmaceutical compositions suitable for topical application to the skin may, for example, take the form of ointments, creams, lotions, pastes, gels, sprays, powders, gels, eye washes, solutions or suspensions, aerosols or oils.

In another embodiment of the invention, a COX-2 inhibitor and LTB₄The composition of the receptor antagonist can be provided in a pharmaceutically acceptable carrier or excipient to form a pharmaceutical composition. Thus, in one embodiment, the invention includes a pharmaceutical composition comprising a COX-2 inhibitor, LTB₄A receptor antagonist and a pharmaceutically acceptable carrier. And, in another embodiment, the invention includes a pharmaceutical composition comprising a COX-2 inhibitor, LTB₄A receptor antagonist and a pharmaceutically acceptable excipient.

Pharmaceutically acceptable carriers and excipients include, but are not limited to, physiological saline, Ringer's solution, phosphate solution or buffer, buffered saline, and other carriers known in the art. The pharmaceutical composition may also include stabilizers, antioxidants, colorants, and diluents. The pharmaceutically acceptable carriers and additives are selected so that side effects of the pharmaceutical compound are minimized and the properties of the compound are not eliminated or inhibited to the extent that the treatment is not effective.

The carrier should be acceptable in the sense of being compatible with the other ingredients of the composition and not deleterious to the recipient thereof. The carrier may be a solid or a liquid or both and is preferably formulated with the compound as a unit dose composition, e.g., a tablet, which may contain from 0.05% to 95% by weight of the active compound.

Carriers that may be used include petrolatum (e.g., Vaseline ), lanolin, polyethylene glycols, alcohols, and combinations of two or more thereof.

Solid dosage forms useful in the methods of the present invention include tablets, capsules, pills, and granules, which may be prepared using enteric coatings and other coatings and shells, for example, as are known in the art.

Compositions for oral administration may be prepared according to any method known to the art for the preparation of pharmaceutical compositions which may contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents, taste masking agents and preserving agents in order to provide pharmaceutically beneficial, palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of pharmaceutical tablets. These excipients may be, for example, inert diluents (such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, sodium phosphate, microcrystalline cellulose or mannitol), granulating and disintegrating agents (for example, corn starch or alginic acid), binding agents (for example, starch, gelatin, gum arabic, hydroxypropyl cellulose, hydroxypropyl methylcellulose or polyvinylpyrrolidone), disintegrating agents (for example, sodium starch glycolate or croscarmellose sodium) and lubricating agents (for example, magnesium stearate, stearic acid or talc). The tablets may also contain a glidant such as silicon dioxide and a wetting agent such as sodium lauryl sulfate. The tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delayed feedstock such as glyceryl monostearate or glyceryl distearate may be employed.

Oral formulations may be presented as hard gel capsules wherein the active ingredient is mixed with an inert solid diluent, for example, lactose, mannitol, microcrystalline cellulose, calcium carbonate, calcium phosphate or kaolin, or as soft gel capsules wherein the active ingredient is present as such or is mixed with water or an oil medium, for example peanut oil, liquid paraffin, soybean oil, olive oil or fractionated coconut oil.

Aqueous suspensions may be prepared containing the active material in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth, xanthan gum and gum acacia. Dispersing or wetting agents may be a naturally occurring phosphatide, for example lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example cetostearyl oxide, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyoxyethylene sorbitan monooleate.

The aqueous suspensions may also contain one or more preservatives, for example ethyl or n-propyl p-hydroxybenzoate, one or more colouring agents, one or more flavouring agents or one or more sweetening agents, for example sucrose or saccharin

Oily suspensions may be formulated by suspending the active ingredient in an omega-3 fatty acid, in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol.

Sweetening agents such as those set forth above, and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an antioxidant such as ascorbic acid.

Dispersible powders and granules suitable for use in the preparation of an aqueous dispersion by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents have been mentioned above. Other excipients, for example sweetening, flavoring and coloring agents, may also be present.

The active ingredient may also be administered as a composition by injection, wherein saline, dextrose or water may be the appropriate carrier.

Suitable inhalable formulations include the active ingredient in a liquid carrier. The carrier is typically water (most preferably sterile, pyrogen-free water) or a dilute aqueous alcohol solution (preferably an artificial isotonic solution or one which is hypertonic for body fluids by the addition of, for example, sodium chloride). Optional additives include preservatives (if the formulation is not sterilized), for example, methyl hydroxybenzoate, and antioxidants, flavoring agents, volatile oils, buffering agents, and surfactants, which are typically used during the preparation of pharmaceutical compositions.

The compositions of the present invention may also be administered rectally. These compositions can be prepared by the following steps: mixing a compound of the invention with one or more suitable non-irritating excipients, for example cocoa butter, synthetic mono-, di-or triglycerides, fatty acids and polyethylene glycols (which excipients are solid at ordinary temperatures but liquid at rectal temperature and will therefore melt in the rectum and release the drug); then, the resulting mixture was molded.

The compositions of the present invention may optionally be supplemented with other agents, such as viscosity enhancers, preservatives, surfactants, and penetration enhancers.

Viscosity is important for some drugs. Droplets with high viscosity tend to stay in the body for longer periods of time and thus increase the absorption of the active compound by the target tissue or increase the retention time. Such viscosity building agents include, for example, polyvinyl alcohol, polyvinyl pyrrolidone, methyl cellulose, hydroxypropyl methylcellulose, hydroxyethyl cellulose, carboxymethyl cellulose, hydroxypropyl cellulose, or other agents known to those skilled in the art. These agents are generally used in amounts of 0.01% to 2% by weight.

Preservatives may optionally be employed to avoid microbial contamination during use. Suitable preservatives include polyquaternium-1, benzalkonium chloride, thimerosal, chlorobutanol, methylparaben, propylparaben, p-hydroxyphenylethyl alcohol, disodium edetate, sorbic acid or other agents known to those skilled in the art. Polyquaternium-1 is preferably used as the antimicrobial preservative. Typically, such preservatives are used in amounts of 0.001 wt% to 1.0 wt%.

The solubility of the components of the compositions of the present invention may be enhanced by surfactants or other suitable co-solvents in the compositions. Such co-solvents include polysorbates 20, 60, and 80, polyoxyethylene/polyoxypropylene surfactants (e.g., Pluronic F-68, F-84, and P-103), cyclodextrins, or other agents known to those skilled in the art. Typically, such co-solvents are used in amounts of 0.01 wt% to 2 wt%.

A penetration enhancer is an agent used to increase the permeability of the skin to an active agent to increase the rate at which the drug diffuses through the skin and into the tissue and bloodstream. Thus, in one embodiment of the present invention, a penetration enhancer may be added to the COX-2 inhibitor and LTB₄In a topical composition of a receptor antagonist.

Examples of penetration enhancers suitable for use with the compositions of the present invention include alcohols, e.g., ethanol and isopropanol; polyols, such as n-alkanols, limonene, terpenes, dioxolanes, propylene glycol, ethylene glycol, other diols, and glycerol; sulfoxides such as dimethyl sulfoxide (DMSO), dimethylformamide, methyldodecyl sulfoxide, dimethylacetamide; esters such as isopropyl myristate/palmitate, ethyl acetate, butyl acetate, methyl propionate and capric/caprylic triglyceride; a ketone; amides, such as acetamide; oleates, such as triolein; various surfactants, such as sodium lauryl sulfate; various alkyl acids, for example, octanoic acid; lactam compounds, for example, laurocapram (azone); alkanols, for example, oleyl alcohol; dialkylaminoacetates and mixtures thereof.

Pharmaceutically acceptable excipients and carriers include all of the foregoing and the like. The above considerations regarding effective formulation and administration procedures are well known in the art and described in standard manuals.

Depending on various factors, dosage regimens are selected to inhibit, ameliorate, or ameliorate the disease conditions described herein. These factors include the type, age, weight, sex, diet and physical condition of the patient, and the severity of the disease, the route of administration, pharmacological considerations such as activity, efficacy, pharmacokinetics and toxicological profiles of the particular compound employed, whether a drug delivery system is employed and whether the compound is administered as part of a pharmaceutical combination. The dosage regimen actually employed may thus vary within wide limits and therefore deviate from the preferred dosage regimen set forth above.

The amount of a composition of the invention required to achieve a desired biological effect will, of course, depend on a variety of factors, such as the particular composition selected, the use of the composition, the mode of administration, and the clinical condition of the patient and recipient to be treated. Patients with the treated disease can initially be treated initially with the dosages indicated above. Treatment should generally last from weeks to months or years until the disease condition has been controlled or eliminated. Patients treated with the compounds or compositions disclosed herein can be routinely monitored to determine the effect of the therapy. Continuous analysis of these data allows for modification of treatment regimens during treatment, resulting in administration of the optimal effective amount of a compound of the invention at any time point and also allows determination of the duration of treatment. In this way, the treatment regimen/dosage schedule can be reasonably modified during the course of treatment so that the lowest amount of LTB is administered₄Receptor antagonists and COX-2 selective inhibitors have satisfactory results and administration is continued only until the time required for successful treatment of the disease.

Pharmaceutical compositions according to the invention include those suitable for oral, rectal, topical, buccal (e.g. sublingual) and parenteral (e.g. subcutaneous, intramuscular, intradermal, intravertebral, intramedullary or intravenous) administration, although the most suitable route in any given case will depend on the nature and severity of the condition being treated and on the nature of the particular compound being adapted. In most cases, the route of administration is oral.

In another embodiment, the present invention provides a kit comprising a container having disposed therein a composition of the present invention.

The oral dosage may be administered as a daily dose or as a single two-day dose or as multiple doses spaced throughout the day. For oral administration, the pharmaceutical composition may be in the form of, for example, tablets, capsules, suspensions or liquids. Capsules, tablets, and the like may be prepared by conventional methods well known in the art. The pharmaceutical compositions are preferably prepared in the form of dosage units containing the active ingredient in the specified amount. Examples of dosage units are tablets or capsules, and may contain one or more therapeutic compounds in the amounts described herein.

The compositions of the present invention may also be administered enterally, by inhalation spray, rectally, topically, buccally or parenterally in the form of dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles as desired. Parenteral administration includes subcutaneous, intramuscular, intradermal, intramammary, intravenous and other methods of administration known in the art. Enteral administration includes solutions, tablets, sustained release capsules, enteric coated capsules and syrups. When administered, the pharmaceutical composition may be generally at or near body temperature.

Oral delivery of the compositions of the present invention may include formulations known in the art which deliver a tablet into the gastrointestinal tract by any of a variety of mechanisms, either delayed or sustained. These mechanisms include, but are not limited to, pH sensitive release from the dosage form based on changes in small intestine pH, slow erosion of tablets or capsules, retention in the stomach based on physical properties of the formulation, bioadhesion of the dosage form to the mucosal lining of the intestinal tract, or enzymatic release of the active drug from the dosage form. For some therapeutic compounds that may be used in the above methods, the expected effect of the combinations and compositions of the present invention is to prolong the time of release of the active drug molecule to the site of action by manipulating the dosage form. Thus, enteric coated formulations and enteric controlled release formulations are within the scope of the invention. Suitable enteric coatings include cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate, and anionic polymers of methacrylic acid and methyl methacrylate.

Pharmaceutical compositions suitable for parenteral administration may generally comprise sterile aqueous preparations of a compound of the invention. These formulations are preferably administered intravenously, but administration can also be effected by subcutaneous, intramuscular or intradermal injection or by infusion. Such formulations are generally prepared by the following steps: the compound is mixed with water and the resulting solution is sterilized and isotonic with blood. The injectable composition according to the invention typically comprises 0.1-10% w/w of the compounds disclosed herein.

Administration can also be delivered intravaginally by use of an intravaginal device. Vaginal delivery may be desirable for some target subjects because, due to absorption from the vagina, more than 10 to 30 times more orally delivered therapeutic agents can be delivered (due to absorption from the vagina), which far exceeds the absorption of drugs by the gastrointestinal tract. Additionally, vaginal administration generally avoids the major problems associated with oral administration, such as gastroesophageal reflux and ulceration.

Transdermal administration is also possible. Pharmaceutical compositions suitable for transdermal administration may be presented as suitable discrete patches to remain in intimate contact with the epidermis of the recipient for an extended period of time. These patches suitably comprise a compound of the invention in an optionally buffered aqueous solution, dissolved and/or dispersed in a binder or dispersed in a polymer. Suitable concentrations of the active compound are about 1% to 35%, more preferably about 3% to 15%. As a particular possibility, the compounds can be prepared, for example, by reaction of the compounds described in Pharmaceutical Research 3 (6): 318(1986) is delivered by the patch described above.

The methods of the invention are useful for, but not limited to, preventing and/or treating pain or inflammation or disorders associated with inflammation. For example, in some embodiments, the disorder associated with inflammation is arthritis. In another example, the compounds described herein may be used to treat pain or inflammation as described below or any inflammation-related disorder, for example as an analgesic in the treatment of pain and headache, or as an antipyretic for the treatment of fever. The compounds described herein will also be useful for treating inflammation-related disorders in a target subject having an inflammation-related disorder.

In the present invention, COX-2 inhibitor and LTB₄The anti-inflammatory effects of each and both inhibitors together can be assessed using the murine Air Pouch model (see example 1 and figure 1). In the treatment of COX-2 inhibitors or LTB₄After addition of the inhibitor or both to the zymosan stimulated Air Pouch, the results can be reported in terms of the number of cells that have permeated in each vessel as an indicator of inflammation.

In some embodiments, the methods and compositions of the present invention comprise preventing and/or treating disorders associated with inflammation. In other embodiments, the methods and compositions of the invention comprise preventing and/or treating any one or more disorders selected from the group consisting of: connective tissue and joint disorders, pain and pain-related disorders, neoplasia disorders, cardiovascular disorders, ear disorders, eye disorders, respiratory disorders, gastrointestinal disorders, angiogenesis-related disorders, immune disorders, allergic disorders, nutritional disorders, infectious diseases and disorders, endocrine disorders, metabolic disorders, neurological and neurodegenerative disorders, psychiatric disorders, liver and bile disorders, musculoskeletal disorders, genitourinary disorders, gynecological and obstetrical disorders, injury and trauma disorders, surgical disorders, dental and oral disorders, sexual dysfunction disorders, skin disorders, blood disorders and toxic disorders.

The invention also includes the treatment of benign, pre-cancerous or malignant neoplasia.

The compositions of the invention are useful in the prevention or treatment of benign and malignant tumors and neoplasia, including cancers such as colon cancer, brain cancer, bone cancer, epithelial cell-derived neoplasia (epithelial cancer) such as basal cell carcinoma, adenocarcinoma, gastrointestinal cancer such as lip cancer, oral cancer, esophageal cancer, small bowel cancer and stomach cancer, colon cancer, liver cancer, bladder cancer, pancreatic cancer, ovarian cancer, cervical cancer, lung cancer, breast cancer and skin cancers such as squamous cell carcinoma and basal cell carcinoma, prostate cancer, renal cell carcinoma and other known cancers that affect epithelial cells throughout the body. In one embodiment, the neoplasia is selected from the group consisting of gastrointestinal cancer, Barrett's esophagus, liver cancer, bladder cancer, pancreatic cancer, ovarian cancer, prostate cancer, cervical cancer, lung cancer, breast cancer, and skin cancers such as squamous cell carcinoma and basal cell carcinoma. The compounds may also be used to treat fibrosis associated with radiation therapy. The method may also treat target subjects with adenomatous polyps, including those with Sporadic Adenomatous Polyposis (SAP) or Familial Adenomatous Polyposis (FAP). In addition, the method may be used to prevent polyps in patients at risk for FAP.

In other embodiments, the methods and compositions of the present invention comprise preventing and treating a neoplastic disorder selected from the group consisting of: lentigo-like melanoma on extremities, actinic keratosis, adenocarcinoma, lymphoid tissue carcinoma (adenoid cystic carcinoma), adenoma, familial adenomatous polyposis, familial polyposis, colon polyps, adenosarcomas, cervical adenosquamous carcinoma, adrenocortical carcinoma, AIDS-related lymphoma, anal carcinoma, astrocytic tumor, Pasteurella adenocarcinoma, basal cell carcinoma, cholangiocarcinoma, bladder carcinoma, brain stem glioma, brain tumor, breast carcinoma, bronchial adenocarcinoma, capillary carcinoma, benign tumor, carcinoma, carcinosarcoma, cavernous (calnus), central nervous system lymphoma, brain astrocytoma, cholangiocarcinoma, chondosarcomas, choroideremia/carcinoma, clear cell carcinoma, skin carcinoma, brain carcinoma, colon carcinoma, colorectal carcinoma, cutaneous T-cell lymphoma, cystadenoma, endoblastoma, endometrial hyperplasia, endometrial carcinosarcoma, endometrioid carcinoma cell sarcoma, interstitial carcinomatoid carcinoma, interstitial carcinoma cell carcinoma, endometrial carcinoma, melanoma, and cervical adenocarcinoma, Ependymal (ependomylal), epithelioid (epithyloid), esophageal cancer, Ewing's sarcoma, extragonadal germ cell tumor, laminar fibroid, focal nodule hyperplasia of the liver, gallbladder cancer, gastrinoma, germ cell tumor, trophoblastic tumor, glioblastoma, glioma, pancreatic hyperglycemia tumor, hemangioblastoma, hemangioma, hepatoadenoma, hepatoadenomatosis, hepatocellular carcinoma, Hodgkin's lymphoma, hypopharyngeal bone cancer, hypothalamic and optic glioma, insulinoma, intraepithelial neoplasia (inteipthelialphenoplasia), intraepithelial squamous cell neoplasia, intraocular melanoma, squamous cell carcinoma, large cell carcinoma, small cell carcinoma, Kaposi's sarcoma, kidney cancer, larynx cancer, leiomyosarcoma, lentigo melanoma, leukemia-related disorders of the oral cavity, lip cancer and cancer, liver cancer, lung cancer, lymphoma, malignant mesothelioma, malignant neuroblastoma, medullary carcinoma of the mouth, medullary carcinoma of the stomach, medullary carcinoma of the eye, and other tumors Melanoma, meninges (meningeal), Merkel cell carcinoma, mesothelial (mesothelial), skin metastatic carcinoma, mucoepidermoid carcinoma, multiple myeloma/plasmacytoma, mycosis fungoides, myelodysplastic syndrome, myeloproliferative and extramedullary dysproliferative disorders, cancer of the nasal cavity and sinuses, nasopharyngeal carcinoma, neuroblastoma, neuroepithelial adenocarcinoma nodular melanoma, non-Hodgkin lymphoma, oat cell carcinoma, oligodendroglia (oligodendroglioma), oral cancer, oropharyngeal cancer, osteosarcoma, pancreatic polypeptide, ovarian cancer, ovarian germ cell tumor, pancreatic cancer, papillary serous adenocarcinoma, pineal gland cell, pituitary tumor, plasmacytoma, pseudosarcoma, lung embryoma, parathyroid cancer, penile cancer, pheochromocytoma, pinebulbar/epineublastic tumor, pituitary tumor, plasmacytoma, pleural cell tumor, prostate cancer, and adrenal gland cell tumor, Rectal cancer, renal cell carcinoma, eye cancer, rhabdomyosarcoma, serous cancer, small cell carcinoma, small bowel cancer, soft tissue cancer, somatostatin-secreting tumor, squamous carcinoma, squamous cell carcinoma, subcutaneous mesothelial (submethlial), superficial diffusible melanoma, supratentorial primitive extraneoembryonic tumor, thyroid cancer, undiffentiated cancer, urinary tract cancer, uterine sarcoma, uveal melanoma, verrucous cancer, leaf tip cancer, VIP tumor, vulvar cancer, Waldenstrom macroglobulinemia, hyper-differentiated cancer, and Wilm tumor.

In another embodiment, the compositions of the present invention are useful for treating symptoms and syndromes resulting from the treatment of cancer. For example, the compositions of the present invention may be used to treat chemotherapy or radiation-induced malignancies.

In another embodiment, the methods and compositions of the invention comprise preventing and treating connective tissue and joint disorders selected from the group consisting of: arthritis, rheumatoid arthritis, spondyloarthopathies, gouty arthritis, degeneration of lumbar joints, carpal tunnel syndrome, canine hip dysplasia, systemic lupus erythematosus, juvenile arthritis, osteoarthritis, tendonitis, and bursitis.

In another embodiment, the methods and compositions of the invention comprise preventing and treating cardiovascular disorders selected from the group consisting of: myocardial ischemia, hypertension, hypotension, cardiac arrhythmias, pulmonary hypertension, hypokalemia, vascular disease, vascular rejection, atherosclerosis including cardiac transplant atherosclerosis, myocardial infarction, embolism, stroke, thrombosis including venous thrombosis, angina including unstable angina, coronary plaque inflammation, cardiac ischemia, myocardial embolism, cardiac remodeling, cardiac fibrosis, myocardial necrosis, aneurysm, arterial fibrosis, embolism, vascular plaque inflammation, vascular plaque rupture, bacterial and viral-induced inflammation, edema, swelling, fluid accumulation, liver cirrhosis, Bartter syndrome, myocarditis, occlusive atherosclerosis, calcification (e.g., arterial and valve calcification), coronary heart disease, heart failure, congestive heart failure, shock, arrhythmia, left ventricular hypertrophy, stroke, vascular graft, arterial thrombosis, myocardial infarction, arterial thrombosis, arterial, Angina, diabetic nephropathy, renal failure, eye injury, migraine, aplastic anemia, heart loss, diabetic cardiomyopathy, renal insufficiency, renal damage, renal arteriopathy, peripheral vascular disease, cognitive dysfunction, stroke, headache, and inflammation associated with surgical procedures, such as vascular grafts (including coronary bypass surgery), revascularization procedures (including angioplasty, stenting, arterial end resection or other invasive procedures involving arteries, veins and capillaries)

In another embodiment, the methods and compositions of the present invention comprise preventing and treating a metabolic disorder selected from the group consisting of: obesity, overweight, type I and type II diabetes, hypothyroidism and hyperthyroidism.

In another embodiment, the methods and compositions of the present invention comprise preventing and treating a respiratory disorder selected from the group consisting of: asthma, bronchiectasis, Chronic Obstructive Pulmonary Disease (COPD), cystic fibrosis, pulmonary edema, pulmonary embolism, pneumonia, pulmonary sarcoidosis (pulmony sarcodosis), silicosis, pulmonary fibrosis, respiratory failure, acute respiratory distress syndrome, and emphysema.

In another embodiment, the methods and compositions of the present invention comprise preventing and treating an angiogenesis-related disorder selected from the group consisting of: angiofibroma, neovascular glaucoma, arteriovenous malformations, arthritis, Osler-Weber syndrome, atherosclerotic plaque, psoriasis, corneal graft neovascularization, suppurative granuloma, delayed wound healing, retrolental fibroplasia, diabetic retinopathy, scleroderma, granulation, solid tumors, hemangioma, trachoma, hemophilous joints, vascular adhesion, hypertrophic scars, age-related macular visual degeneration, coronary heart disease, stroke, cancer, AIDS complications, ulcers and infertility.

In another embodiment, the methods and compositions of the invention include the prevention and treatment of a susceptible disease or disorder selected from the group consisting of: viral infection, bacterial infection, Prion infection, helicobacter infection, mycobacterial infection, Rickettsia infection, chlamydia infection, parasitic infection and fungal infection.

In another embodiment, the methods and compositions of the invention comprise the prevention and treatment of infectious diseases and disorders selected from the group consisting of: hepatitis, hiv (aids), smallpox, chicken pox, cold, bacterial influenza, viral influenza, warts, oral herpes, genital herpes, herpes simplex infection, herpes zoster, bovine spongiform encephalopathy, septicemia, streptococcal infection, staphylococcal infection, anthrax, Severe Acute Respiratory Syndrome (SARS), malaria, african narcolepsy, yellow fever, chlamydia infection, botulism, canine heartworm disease, rocky mountain spotted fever, lyme disease, cholera, syphilis, gonorrhea, encephalitis, pneumonia, conjunctivitis, fungal infection, rabies, dengue fever, ebola, measles, mumps, rubella, West Nile virus, meningitis, gastroenteritis, tuberculosis, hepatitis, and scarlet fever.

The invention also provides compositions comprising COX-2 inhibitors and LTB₄A therapy of a receptor antagonist combination, said therapy comprising the treatment and prevention of a neurodegenerative disorder in a patient suffering from: for example, dementia, aphasia, amnesia, depression, psychogenic apraxia, anxiety, pathologic personality, cognition deficit, and hallucinations.

The term "neurodegenerative disorder" as used herein is defined as any abnormality in one or more of the root nerves, a postoperative disease of any tissue comprised of nerves or an age-related disease of one or more of the root nerves. The term "nerve" as used herein includes any composition or result found in or on the central or peripheral nervous system, including, but not limited to, nerve cells, brain tissue, spinal cord tissue, glial cells, astrocytes, dendrites, choline receptors, adrenergic receptors, gamma aminobutyric acid receptors, serotonin-activated (5-HT) receptors, glutamate receptors, endorphin-enkephalin (opioids) receptors, Schwann cells, axons, oligodendrocytes, microglia, ependymal membranes, myelin sheaths, and any other neurological tissue in the body of a target subject.

The term "neurodegenerative disorder" also includes any complication arising from having the above-mentioned disorder. For example, some chronic neurodegenerative disorders are often associated with complications such as those caused by immobility, muscle contractures, shortened lifespan, opportunistic infections, and stress, and any complications that may ultimately result from chronic or recurrent neurodegenerative disorders. Behavioral neurodegenerative disorder complications include resistance, aggression, agitation, absentmindedness, and absence of cooperation. Psychiatric complications include depression, anxiety, paranoid reactions, delusions and hallucinations.

Neurodegenerative disorders can arise in a target subject via several determinants, including chronic substance abuse, vascular disease and inappropriate consumption of vitamins, invasive agents, pathogenic agents, brain cancer, mental or physical trauma, brain trauma, and genetics. The methods and compositions of the invention are intended to treat a target subject suffering from a neurodegenerative disorder, regardless of how the disorder is first caused.

In one embodiment, the methods and compositions of the present invention comprise preventing and treating neurodegenerative disorders selected from the group consisting of: cortical dementia, dementia of general, aging, Alzheimer's disease, vascular dementia, multiple dementia, pre-senile dementia, alcohol-induced dementia, senile dementia, stroke, coma, seizure (seizure), epilepsy, amnesia, hypovolemic shock, phenylketonuria, aminonuria, Tay-Sachs disease, Niemann-Pick, Gaucher's disease, Hurler's syndrome, Krabbe's disease, leukodystrophy, traumatic shock, reperfusion injury, multiple sclerosis, AIDS-related dementia, neurocytotoxicity, brain trauma, adult respiratory disease (Creards), acute spinal cord injury (acute spinal cord injure), Parkinson's disease, frontotemporal dementia, Pick's disease, ischemia, paralysis, supranuclear palsy, cortical basal distortion, multiple dementia, mental-koutzfeldt-kob disease, Jazheimer's disease, normal pressure hydrocephalus, parakinson, confusion disorder, hydrocephalus, amnesia, hydrocephalus, dementia, Alzheimer's disease, Parkinson's disease, dementia, Alzheimer Aging, muscular dystrophy, ALS, muscular dystrophy, epilepsy, schizophrenia, depression, anxiety, autism, phobias, spongiform encephalopathy, Huntington's chorea, ischemia, obsessive-compulsive disorder, anxiety-related disorders, stress-related disorders, psychosis, neuroendocrine system disorders, thermoregulatory disorders, vasoactive headaches, sexual dysfunction, tooth morphogenetic disorders, Tourette's syndrome, autism, attention deficit disorders, hyperactivity disorders, sleep disorders, social phobias, urinary incontinence, vasospasm, stroke, eating disorders such as obesity, anorexia, and bulimia, binge depression, bipolar disorder, drug addiction, alcoholism, and tobacco addiction. In addition, neurodegenerative disorders that can be treated using the compositions and methods described herein include target subjects that are otherwise normal, but for which it is desirable to improve certain cognitive abilities, such as memory retention and thought processes.

In another embodiment, the methods and compositions of the present invention comprise preventing and treating a skin disorder selected from the group consisting of: acne, psoriasis, eczema, burns, poison ivy, poison oak, and dermatitis.

In another embodiment, the methods and compositions of the present invention comprise preventing and treating a surgical disorder selected from the group consisting of: post-operative pain and swelling, post-operative infection and post-operative inflammation.

In another embodiment, the methods and compositions of the present invention include the prevention and treatment of gastrointestinal disorders, including the group consisting of: enteritis, irritable bowel syndrome, Crohn's disease, gastritis, irritable bowel syndrome, diarrhea, constipation, dysentery, ulcerative colitis, gastroesophageal reflux, gastric ulcer, gastric varices, ulcers and heartache.

In another embodiment, the methods and compositions of the present invention comprise preventing and treating an otic disorder selected from the group consisting of: ear pain (otic pain), inflammation, otorrhea, ear pain, fever, ear bleeding, Lermoyez syndrome, Meniere's disease, vestibular neuronitis, benign paroxysmal positional vertigo, herpes zoster of the ear, Ramsay Hunt syndrome, viral neuronitis, gangliitis, herpes benomyeli, otitis interna, purulent otitis interna, viral endolymphatic otitis, perilymphatic fistula, noise-induced hearing loss, presbycusis, drug-induced ototoxicity, acoustic neuroma, barotropic otitis media, infectious myringitis, giant myringitis, otitis media with effusion, acute otitis media, secretory otitis media, acute generalized papillottitis, chronic otitis media, otitis extema, otosclerosis, squamous cell carcinoma, basal cell carcinoma, non-chromophoric sarcoidosis, chemoreceptor tumor, Jugular venous spheroids (globus jugulatus), tympanostomy, otitis externa, pouchitis, eczematoid dermatitis, malignant otitis externa, subperiondronic hematoma, cerominomas, cerumen embolism, sebaceous cysts, osteoma, keloids, ear pain, tinnitus, vertigo, tympanic membrane infection, typanitis, ear furuncles, otorrhea, acute mastoplasia, petroselitis, conductive and sensorineural hearing loss, spinal epidural abscess, lateral sinus thrombosis, subdural pyogenesis, otitis hydrocephalus, Dandy syndrome, bullous myringitis, cerumen embolism, diffuse otitis externa, foreign body reaction, obstructive keratosis, otoma, otomycosis, trauma, acute barotrauma, acute eustachian tube occlusion, post-otic surgery (post-otoicsurgery), post-operative earache, pearloma, conductive and sensorineural hearing loss, spinal epidural abscess, lateral sinus thrombosis, subdural pyogenesis, and otitis hydrocephalus.

In another embodiment, the methods and compositions of the present invention comprise preventing and treating an ocular disorder selected from the group consisting of: retinopathy, uveal eye, photophobia, acute loss to eye tissue, conjunctivitis, age-related macular degeneration diabetic retinopathy, retinal detachment, glaucoma, vitelliform macular dystrophy, choroidal and retinal swirl atrophy, conjunctivitis, corneal infections, Fuch dystrophy, iridocorneal endothelial syndrome, retinitis, keratoconus, lattice degeneration, corneal epithelial basement membrane dystrophy, ocular herpes, pterygium, myopia, hyperopia, and cataracts.

The compositions and methods of the invention may also be used in the treatment of pain, but are not limited to, post-operative pain, dental pain, muscle pain, neuropathic pain, and pain caused by cancer.

In another embodiment, the methods and compositions of the present invention comprise preventing and treating dysmenorrhea, kidney stones, soft tissue injuries, wound healing, vaginitis, candidiasis, sinus headache, pressure headache, periarteritis nodosa, thyroiditis, myasthenia gravis, sarcoidosis, nephrotic syndrome, Bahcet syndrome, polymyositis, gingivitis, hypersensitivity reactions, post-traumatic swelling, closed head injury, liver disease, and endometriosis.

The methods and compositions of the invention encompass not only the prevention or treatment of pain or inflammation-related disorders in humans, but also the prevention or treatment of such disorders in various animals. For example, some animals may also suffer from adverse consequences associated with pain or inflammation or disorders associated with inflammation. Moreover, some disorders associated with inflammation respond in dogs to the same treatments as those used in humans. Thus, in addition to being useful in humans, the methods and compositions of the present invention also include the treatment and prevention of pain or inflammation and in some cases inflammation-related disorders in other mammals, including: horses, dogs, cats, sheep, pigs, cattle, etc. Thus, preferably, the target object is an animal, more preferably, the target object is a mammal. Preferably, the mammal is a human.

It will be appreciated that the amount of the composition of the invention used in the treatment or prevention of the diseases described herein will vary within wide limits, being adjusted to the needs of the individual in each particular case. In general, for administration to adults, suitable daily dosages are described herein, but the dosages identified herein may be in excess if advantageous. The daily dose may be administered as a single dose or in divided doses.

Suitable dosage levels of COX-2 inhibitor are generally in the range of about 0.01mg/kg to about 140mg/kg of the body weight of the subject of interest per day, and the inhibitor may be administered in a single or multiple doses. In one embodiment, the dosage level is from about 0.1mg/kg to about 25mg/kg per day, and in another embodiment, from about 0.5mg/kg to about 10mg/kg per day.

In larger mammals, such as humans, a dosage of about 0.5mg to 7 grams per day is generally required for oral administration. The compounds may be administered several times daily, for example, 1-4 times daily, or 1 or 2 times daily.

The amount of COX-2 inhibitor that can be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. For example, a formulation for oral administration to humans may contain 0.5mg-7g of the active agent, optionally mixed with an appropriate and convenient amount of a carrier material which may vary from about 5 to about 95% of the total composition. For COX-2 inhibitors, unit dosage forms typically contain from about 1mg to about 500mg of the active ingredient, typically 25mg, 50mg, 100mg, 200mg, 300mg, 400mg, 500mg, 600mg, 800mg, or 1000 mg.

The total daily dose of the COX-2 inhibitor may be generally in the range of about 0.001 to about 10000 mg/day (in single or divided doses), and in one embodiment in the range of about 1.0mg to about 2000 mg. However, it will be understood that the specific dosage level of a therapeutic agent or method of treatment of the present invention for any particular patient will depend upon a variety of factors including: the activity of the particular compound employed, the age, body weight, general health, sex and dietary profile of the patient, the time of administration, the rate of excretion, the composition of the drug and the severity of the particular disease to be treated and the form of administration.

In one embodiment of the invention, LTB₄The dosage of the compound is from about 0.01mg to about 5000mg or any other dosage according to the particular regulator.

In one embodiment of the invention, a COX-2 inhibitor is combined with LTB₄The ratio of receptor antagonists was 1: 1. In another embodiment of the invention, a COX-2 inhibitor and LTB₄The ratio of receptor antagonists is any concentration from 1 to greater than 1. For example, in one embodiment, a COX-2 inhibitor is combined with LTB₄The ratio of receptor antagonists was 1: 2. In bookIn another embodiment of the invention, a COX-2 inhibitor and LTB₄The ratio of receptor antagonists is any concentration ratio of 1 greater than 1. For example, in one embodiment of the invention, a COX-2 inhibitor is combined with LTB₄The ratio of receptor antagonists was 2: 1. For example, in one embodiment, the composition comprises about 200mg celecoxib and about 200mg LTB₄A receptor antagonist. In another embodiment of the invention, the composition comprises about 200mg celecoxib and about 400mg LTB₄A receptor antagonist, in another embodiment of the invention, the composition comprises about 100mg celecoxib and about 200mg LTB₄A receptor antagonist.

Therapeutic doses are usually titrated to optimize safety and efficacy. Generally, an initial dose-effect relationship in vitro can provide beneficial guidance for the appropriate dose to be administered to a patient. Studies in animal models can also be used for guidance regarding effective dosages for treating pain or inflammation according to the present invention in general. Depending on the treatment regimen, it will be understood that the dosage form to be administered will depend on a variety of factors including the particular agent being administered, the route of administration and the condition of the particular patient. In general, it is desirable to administer an amount of the compound that is effective to achieve plasma levels comparable to the effective concentrations found in vitro. Thus, where the in vitro activity of a compound is found to be, for example, 10 micromolar, it is desirable to administer an amount of drug to provide a concentration of preferably about 10 micromolar in vivo. The determination of these parameters is known to the person skilled in the art.

For the combination therapies provided herein, the dosage can be determined and adjusted based on the therapeutic effect shown in reducing or preventing the symptoms of pain or inflammation-related disorder. In addition, any person of ordinary skill in the art knows how to measure and quantify pain or inflammation symptoms or lack thereof.

c. Detailed description of the preferred embodiments

The starting materials for use in the compositions and methods of the invention are known or may be prepared by conventional methods known to those skilled in the art or in a similar manner to those described in the art.

In general, the process of the invention can be carried out as follows.

The compounds used in the following examples are indicated herein by their example numbers (see tables 1 and 2). For example, LTB in Table 1₄The compound L-1 is 2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl]-4- (trifluoromethyl) benzoic acid. COX-2 inhibitors such as C-1 (celecoxib) can be found in Table 2.

Example 1 COX-2 inhibitor Activity and in vitro COX-1 Selectivity

The in vitro COX-2 inhibitory activity of the compounds as illustrated in the above examples was determined by the following method. The COX-2 inhibitory activity of other COX-2 inhibitors of the invention may also be determined by the following method.

Step 1: preparation of recombinants of COX baculovirus (baculovir)

COX-1 and COX-2 recombinants were determined according to Gierse et al J.biochem.305: 479-84 (1995). A2.0 kb fragment containing the human or murine COX-1 or human or murine COX-2 coding region was cloned into the BamH1 site of the Baculovirus transfer vector pVL1393(Invitrogen) to hybridize with O' Reilly et al in Bacillus Expression Vectors: the baculovirus transfer vectors for COX-1 and COX-2 were generated in a similar manner to the method described in the Arabidopsis Manual (1992). Isolation of baculovirus recombinants by the following method: 4. mu.g of baculovirus transfer vector DNA was infected by calcium phosphate method into SF9 insect cells (2X 108) with 200ng of linearized baculovirus plasmid DNA. See A Manual of Methods for bacterial cells and Instrument Procedures, Texas standard. Exp.Station Bull.1555(1987) by Summer et al. The recombinant virus was esterified by three rounds of spot purification and high titer (107-108 pfu/mL) of the virus material was prepared. For large scale production, SF9 insect cells were infected with recombinant baculovirus stock in a 10 liter fermentor (0.5X 106/mL) to give an infection diversity (multiplicity of infection) of 0.1. After 72 hours, the cells were centrifuged and the cell pellet was homogenized in Tris/Sucrose (50 mM: 25%, pH8.0) containing 1% 3- [ (3-cholineaminopropyl) -dimethylamino ] -1-propanesulfonate (CHAPS). The homogenate was centrifuged at 10000 XG for 30 minutes and the resulting supernatant was stored at-80 ℃ before being assayed for COX activity.

Step 2: COX-1 and COX-2 activity assays

Detection of released prostaglandins by ELISA COX activity was tested according to the PGE2 formed/micro protein/time. CHAPS-lysed insect cell membranes containing the appropriate COX enzyme were incubated in potassium phosphate buffer (50mM, pH8.0) containing epinephrine, phenol, and ferrochromine and arachidonic acid (10 micromolar) was added. The compounds were pre-incubated with enzyme for 10-20 minutes before arachidonic acid was added. After 10 minutes, any reaction between arachidonic acid and the enzyme was stopped at 37 ℃/room temperature by transferring 40 microliters of the reaction mixture into 160 microliters of ELISA buffer and 25 micromolar indomethacin. The PGE2 formed was measured by standard ELISA techniques (Cayman Chemical).

And step 3: rapid assay for COX-1 and COX-2 activity

Detection of released prostaglandins by ELISA COX activity was tested according to the PGE2 formed/micro protein/time. CHAPS-lysed insect cell membranes containing the appropriate COX enzyme were incubated in potassium phosphate buffer (0.05M potassium phosphate, pH7.5, 2. mu. mol phenol, 1. mu. mol heme, 300. mu. mol epinephrine) and 20. mu.l of 100. mu. mol arachidonic acid (10. mu. mol) was added. The compounds were pre-incubated with enzyme at 25 ℃ for 10 minutes before addition of arachidonic acid. After 2 minutes, any reaction between arachidonic acid and the enzyme was stopped at 37 ℃/room temperature by transferring 40 microliters of the reaction mixture into 160 microliters of ELISA buffer and 25 micromolar indomethacin. The PGE2 formed was measured by standard ELISA techniques (cayman chemical).

Example 2 LTB₄ReceptorsAntagonist Activity

LTB of Compounds of the invention₄Activity can be determined by comparing the compounds of the invention with radiation-labelled LTB₄Specific LTB on guinea pig spleen membranes₄The ability of the receptor sites to compete. Pelargonium mouse spleen membranes were prepared as described by Chang et al (J. Pharmacology and Experimental Therapeutics 232: 80, 1985).³H-LTB₄Binding assays in the assay contained 50mM TrispH7.3, 10 mM MgCl₂9% methanol, 0.7nM³H-LTB₄(NEN, approx. 200Ci/mmol) and 0.33mg/mg of guinea pig spleen membrane at 150 mg/l. The unlabeled LTB₄At a concentration of 5 micromolar to determine nonspecific binding. Test compounds were added at varying concentrations to evaluate their pairing³H-LTB₄The effect of the combination. The reaction was incubated at 4 ℃ for 30 minutes. Will be combined with³H-LTB₄The membranes were collected by filtration through a glass fiber filter and the amount of binding was determined by luminescence counting. For the test compound, IC₅₀Is 50% inhibition of specificity³H-LTB₄The concentration of binding.

COX-2 inhibitors and LTB₄Combination therapy with receptor antagonists can be assessed as described in the following tests.

Example 3 Induction and assessment of collagen-induced arthritis in mice

A. Inducing collagen-induced arthritis

Arthritis was induced in 8-12 week old male DBA/1 mice by injection of 50 micrograms of type II collagen in total Freunds adjuvant (Sigma) on day 0, at the base of the tail, as described by j.stuart in Annual rev. Compounds were prepared as suspensions in 0.5% methylcellulose (Sigma, st. louis, MO), 0.025% Tween 20 (Sigma). COX-2 inhibitors (examples 1 and 2) and LTB₄The receptor antagonist (example 3) is administered alone, or the COX-2 inhibitor and LTB₄The receptor antagonist is administered in combination. In the injection of glue to non-arthritic animals0.1mL of compound was administered by gavage starting 20 days after the starting material and continuing daily until final assessment on day 55. These animals were then evaluated for the incidence and severity of arthritis several times a week until day 56. Any animals with paw redness or swelling were counted as arthritis. Severity scoring was performed using a 0-3 score for each paw (up to 12/mouse) as described by p.wooley et al, trans. proc., 15, 180 (1983). For each animal in which arthritis was observed, the incidence and severity of arthritis was measured. The incidence of arthritis was tested at a gross level by observing swelling or redness in the paw or toe. Severity was measured according to the following guidelines. Briefly, animals with four normal paws (i.e., no redness or swelling) were scored as 0. Any redness or swelling of the toes or paws was scored as 1. The overall paw swelling or malformation was scored as 2. Joint stiffness was scored as 3.

B. Tissue experiments of paw

To verify the overall assay for non-arthritic animals, tissue experiments were performed. The paws of the slaughtered animals were removed, fixed and limed as previously described at the end of the experiment (r. jonsson j. immunol. methods, 88, 109 (1986)). Samples were paraffin embedded, sectioned and stained with hematoxylin and eosin by standard methods. Stained sections were examined for cellular infiltration, synovial hyperplasia, and, bone and cartilage erosion.

C. Animal dosage range

Animals were dosed with one of the following dose ranges:

4- [5- (4-chlorophenyl) -3- (trifluoromethyl) -1H-pyrazol-1-yl ] benzenesulfonamide at about 3 mpk/day; 7- [3- [2- (cyclopropylmethyl) -3-methoxy-4- [ (methylamino) carbonyl ] phenoxy ] propoxy ] -3, 4-dihydro-8-propyl-2H-1-benzopyran-2-propionic acid at about 3 mpk/day;

4- [5- (3-fluoro-4-methoxyphenyl) -3- (difluoromethyl) -1H-pyrazol-1-yl ] benzenesulfonamide, at about 30 mpk/day; 7- [3- [2- (cyclopropylmethyl) -3-methoxy-4- [ (methylamino) carbonyl ] phenoxy ] -propoxy ] -3, 4-dihydro-8-propyl-2H-1-benzopyran-2-propionic acid at about 10 mpk/day;

4- [5- (3-fluoro-4-methoxyphenyl) -3- (difluoromethyl) -1H-pyrazol-1-yl ] benzenesulfonamide, at about 10 mpk/day; 7- [3- [2- (cyclopropylmethyl) -3-methoxy-4- [ (methylamino) carbonyl ] phenoxy ] -propoxy ] -3, 4-dihydro-8-propyl-2H-1-benzopyran-2-propionic acid at about 10 mpk/day;

4- [5- (4-chlorophenyl) -3- (trifluoromethyl) -1H-pyrazol-1-yl ] benzenesulfonamide, monday, wednesday, friday, at about 10 mpk/day; 7- [3- [2- (cyclopropylmethyl) -3-methoxy-4- [ (methylamino) carbonyl ] phenoxy ] -propoxy ] -3, 4-dihydro-8-propyl-2H-1-benzopyran-2-propionic acid at about 10 mpk/day.

7- [3- [2- (cyclopropylmethyl) -3-methoxy-4- [ (methylamino) carbonyl ] -phenoxy ] -propoxy ] -3, 4-dihydro-8-propyl-2H-1-benzopyran-2-propionic acid is prepared as described in US patent 5310951, which is incorporated herein by reference.

4- [5- (4-chlorophenyl) -3- (trifluoromethyl) -1H-pyrazol-1-yl ] benzene-sulfonamide was prepared as described in US patent 5466823, which is incorporated herein by reference.

4- [5- (3-fluoro-4-methoxyphenyl) -3- (difluoromethyl) -1H-pyrazol-1-yl ] benzenesulfonamide is prepared as described in U.S. Pat. No. 5466823, which is incorporated herein by reference.

D. Collagen-induced arthritis experiments:

male DBA/1 mice at 8-12 weeks were injected at the base of their tail with 50 micrograms of chicken type II collagen in CFA. After 21 days, the animals were supplemented with 50 μ g of chicken type II collagen in IFA. Vehicle (0.5% methylcellulose + 0.025% Tween 80) or COX-2 inhibitor (Compound C-1, 15mpk, bid) or LTB was used starting on day 21₄Receptor antagonists (Compound L-2, 150mpk, bid) or COX-2 inhibitors with LTB₄The combination of receptor antagonists is used to treat animals. On day 56, animals were evaluated for arthritis. On the pawAnimals with inflammation were classified as positive for incidence. In addition, paw swelling was graded according to the standard scale 0-3/paw (total score 12/animal). Table 4 (corresponding to fig. 2) shows the results of this experiment. Two LTB in combination with Compound C-1 in a collagen-induced arthritis model₄Receptor antagonists (Compound L-2 and Compound L-1) were compared. Table 5 (corresponding to fig. 3) shows the results of this study.

Table 6 (corresponding to fig. 4) shows the results of the respective inhibitors throughout the experiment.

TABLE 4

Group of	% incidence	Severity degree	SEM	Weight (D)	SEM
						Normality	0	0	0	27.94	0.728

Media	100	4.8	0.611	24.06	0.417
						COX-2 inhibitors	100	5.1	0.795	24.66	0.433
LTB4ra	90	2.7	0.559	26.77	0.574
						COX-2 inhibitor + LTB4ra	20	0.333	0.238	26.33	0.484

Standard error of SEM-mean

TABLE 5

	% incidence
					Group of	Media	LTB4ra (100mpk) + COX-2 inhibitor (30mpk)	LTB4ra (300mpk) + COX-2 inhibitor (60mpk)	No Rx
21	0	0	0	0
					35	38	20	0	90
44	75	20	0	100
					49	88	40	29	100
56	88	40	29	100

TABLE 6

Group of	% incidence	Severity degree	SEM	Weight (D)	SEM
						Normality	0	0	0	27.94	0.728
Media	100	4.8	0.611	24.06	0.417
						COX-2 inhibitors	100	5.1	0.795	24.66	0.433
LTB4ra	90	2.7	0.559	26.77	0.574
						COX-2 inhibitor + LTB4ra	20	0.333	0.238	26.33	0.484

Standard error of SEM-mean

Example 4

Preparation of a formulation consisting of 700mg COX-2 inhibitor and 700mg LTB₄A receptor antagonist.

Example 5

A formulation was prepared consisting of 350mg of 4- [5- (3-fluoro-4-methoxyphenyl) -3- (difluoromethyl) -1H-pyrazol-1-yl ] benzenesulfonamide and 350mg of 7- [3- [2- (cyclopropylmethyl) -3-methoxy-4- [ (methylamino) -carbonyl ] phenoxy ] propoxy ] -3, 4-dihydro-8-propyl-2H-1-benzopyran-2-propionic acid.

EXAMPLE 6 solution "A" formulation of Compound L-1

Solution formulations were prepared for oral delivery of compound L-1. The 25mg/mL formulation was prepared by the following method: l-1 was first added to deionized water, followed by NaOH to adjust the pH to 7.0. Polyvinylpyrrolidone having an average molecular weight of 10000, 2% (W/W), was added to the solution as a polymerization precipitation inhibitor to reduce the possibility of precipitation in a lower pH environment in the GI tract during oral administration. The above solutions were used to prepare C-1 suspensions for co-administration of two drugs: a formulation of 200mg/mL C-1 suspension in 25mg/mL L-1 solution was prepared by the following method: a25 mg/mL L-1 solution was prepared first, followed by the above procedure. The C-1 solid drug was added and homogenized to ensure uniform particle size.

Example 7 "B" formulation of C-1

The formulation of a 200mg/mL suspension of C-1 in a 25mg/mL solution of 2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl ] -4- (trifluoromethyl) -benzoic acid was prepared by: l-1 was first added to deionized water, followed by NaOH to adjust the pH to 7.0. 2% (W/W) hydroxymethylcellulose (viscosity of 40-60 centipoise in a 2% aqueous solution at 20 ℃) was dissolved in the solution with 5% (W/W) Tween 80. Compound C-1 solid drug was added and homogenized to ensure uniform particle size.

Example 8: solution "C" formulation of Compound L-1

A formulation of 200mg/mL compound C-1 suspension in 25mg/mL L-1 solution was prepared by the following method: l-1 was first added to deionized water, followed by NaOH to adjust the pH to 7.0. 2% (W/W) PEG 400 was added followed by 2% (W/W) hydroxymethylcellulose (2% aqueous solution at 20 ℃ with a viscosity of 40-60 centipoise). Compound C-1 solid drug was added and homogenized to ensure uniform particle size.

Example 9: single layer tablet having two drug substances granulated together

Compound L-1/Compound C-1 dose	200/200mg			200/100mg
							Composition (I)	％w/w	mg/tablet	g/1000 tablets	％w/w	mg/tablet	g/1000 tablets
Compound (I)aL-1	28.57	200.00	200.00	33.33	200.00	200.00
							Compound C-1	28.57	200.00	200.00	16.67	100.00	100.00
b，cLactose	23.36	163.50	163.50	30.50	183.00	183.00
							Sodium Lauryl Sulfate (SLS)	3.00	21.00	21.00	3.00	18.00	18.00
cPolyvinylpyrrolidone (PVP)	2.50	17.50	17.50	2.50	15.00	15.00
							dCroscarmellose sodium	3.00	21.00	21.00	3.00	18.00	18.00
cMicrocrystalline cellulose	10.00	70.00	70.00	10.00	60.00	60.00
							Magnesium stearate	1.00	7.00	7.00	1.00	6.00	6.00
Total amount of	100.00	700.00	700.00	100.00	600.00	600.00

a. Compound C-1 can be used in the form of the free acid or salt; if a salt is used, the total weight of the tablet is increased or the amount of filler (e.g., lactose) is adjusted to accommodate the weight of the counter ion.

b. Lactose may be replaced by other water-soluble excipients, such as mannitol.

c. The polyvinylpyrrolidone may be replaced by other suitable binders, such as hydroxypropyl cellulose or hydroxypropylmethyl cellulose, and the amount of microcrystalline cellulose may be varied to adjust the drug release profile.

d. Croscarmellose sodium may be replaced by other disintegrants (such as sodium starch glycolate or cross-linked polyvinylpyrrolidone) or removed for controlled release of both drugs as required for tablets.

Compound L-1, compound C-1, lactose, SLS, PVP, croscarmellose sodium (partially or fully) and microcrystalline cellulose are mixed together and granulated using a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled and mixed with part or all of the croscarmellose sodium, followed by final mixing with magnesium stearate. The final blend is then compressed into tablets of appropriate size to obtain the desired dosage.

Two tablets containing 200mg of Compound L-1 and 100mg of Compound C-1 were taken at a dose of 400mg of Compound C-1 and 200mg of Compound C-1.

Example 10: bilayer tablet (Compound L-1 controlled Release/Compound C-1 immediate Release)

Compound L-1(200mg) layer				Compound C-1(200mg) layer
								Composition (I)	％w/w	mg/tablet	g/1000 tablets	Composition (I)	％w/w	mg/tablet	g/1000 tablets
Compound (I)aL-1	33.33	200.00	200.00	Compound C-1	40.00	200.00	200.00
								dPolyethylene oxide	30.66	183.94	183.94	b，cLactose	40.75	203.75	203.75
bLactose	17.00	102.00	102.00	Sodium lauryl sulfate	3.00	15.00	15.00
								Sodium chloride	17.00	102.00	102.00	cPolyvinylpyrrolidone (PVP)	2.50	12.50	12.50

Butylated hydroxytoluene	0.01	0.06	0.06	eCroscarmellose sodium	3.00	15.00	15.00
								Silicon dioxide	0.50	3.00	3.00	cMicrocrystalline cellulose	10.00	50.00	50.00
Magnesium stearate	1.50	9.00	9.00	Magnesium stearate	0.75	3.75	3.75
								Total amount of	100.00	600.00	600.00	Total amount of	100.00	500.00	500.00

a. Compound L-1 can be used in the form of the free acid or salt; if a salt is used, the total weight of the tablet is increased or the amount of filler (e.g., lactose) is adjusted to accommodate the weight of the counter ion.

b. Lactose may be replaced by other water-soluble excipients, such as mannitol, polyethylene glycol or sodium chloride.

c. The polyvinylpyrrolidone may be replaced by other suitable binders, such as hydroxypropyl cellulose or hydroxypropylmethyl cellulose, and the amount of microcrystalline cellulose and lactose may be varied to adjust the drug release profile.

d. The polyoxyethylene may be a mixture of more than one molecule with a molecular weight of 200000-5 million.

e. Croscarmellose sodium may be replaced by other disintegrants such as sodium starch glycolate or cross-linked polyvinylpyrrolidone.

The components of both layers were granulated separately.

For the compound L-1 layer, all ingredients except for silicon dioxide and magnesium stearate were granulated using a dry granulation process or a wet granulation process. The granules are dried (if wet granulated), milled and mixed with silicon dioxide, followed by final mixing with magnesium stearate.

For compound C-1 layer, all ingredients except partially crosslinked hydroxymethyl cellulose sodium and magnesium stearate were mixed together and granulated using either a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled and mixed with part or all of the croscarmellose sodium, followed by final mixing with magnesium stearate.

The two final blends are then compressed into bilayer tablets of appropriate size to obtain the desired dosage.

For a dose of 200mg of Compound L-1 and 200mg of Compound C-1, a bilayer tablet having a total weight of 1100mg (comprising 600mg of Compound L-1 granules and 500mg of Compound C-1 granules) was prepared.

For a dose of 100mg of Compound L-1 and 100mg of Compound C-1, bilayer tablets were prepared with a total weight of 550mg (comprising 300mg of Compound L-1 granules and 250mg of Compound C-1 granules).

Example 11: bilayer tablet (Compound L-1 controlled Release/Compound C-1 immediate Release)

Compound L-1(200mg) layer				Compound C-1(100mg) layer
								Composition (I)	％w/w	mg/tablet	g/1000 tablets	Composition (I)	％w/w	mg/tablet	g/1000 tablets
Compound (I)aL-1	33.33	200.00	200.00	Compound C-1	25.00	100.00	100.00
								dPolyethylene oxide	30.66	183.94	183.94	b，cLactose	53.25	213.00	213.00
Lactoseb	17.00	102.00	102.00	Sodium lauryl sulfate	3.00	12.00	12.00
								Sodium chloride	17.00	102.00	102.00	cPolyvinylpyrrolidone (PVP)	2.50	10.00	10.00
Butylated hydroxytoluene	0.01	0.06	0.06	eCroscarmellose sodium	3.00	12.00	12.00
								Silicon dioxide	0.50	3.00	3.00	cMicrocrystalline cellulose	12.50	50.00	50.00
Magnesium stearate	1.50	9.00	9.00	Magnesium stearate	0.75	3.00	3.00
								Total amount of	100.00	600.00	600.00	Total amount of	100.00	400.00	400.00

a. Compound L-1 can be used in the form of the free acid or salt; if a salt is used, the total weight of the tablet is increased or the amount of filler (e.g., lactose) is adjusted to accommodate the weight of the counter ion.

b. Lactose may be replaced by other water-soluble excipients, such as mannitol, polyethylene glycol or sodium chloride.

c. The polyvinylpyrrolidone may be replaced by other suitable binders, such as hydroxypropyl cellulose or hydroxypropylmethyl cellulose, and the amount of microcrystalline cellulose and lactose may be varied to adjust the drug release profile.

d. The polyoxyethylene may be a mixture of more than one molecule with a molecular weight of 200000-5 million.

e. Croscarmellose sodium may be replaced by other disintegrants such as sodium starch glycolate or cross-linked polyvinylpyrrolidone.

The components of both layers were granulated separately.

For LTB₄The compound L-1 layer, all ingredients except for silicon dioxide and magnesium stearate, was granulated by a dry granulation process or a wet granulation process. The granules are dried (if wet granulated), milled and mixed with silicon dioxide, followed by final mixing with magnesium stearate.

For the C-1 layer of the COX-2 inhibitor, all ingredients except the partially cross-linked sodium carboxymethylcellulose and magnesium stearate are mixed together and granulated using a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled and mixed with part or all of the croscarmellose sodium, followed by final mixing with magnesium stearate.

The two final blends are then compressed into bilayer tablets of appropriate size to obtain the desired dosage.

For a dose of 200mg of Compound L-1 and 100mg of Compound C-1, bilayer tablets were prepared with a total weight of 1000mg (comprising 600mg of Compound L-1 granules and 400mg of Compound C-1 granules).

Two tablets were taken to give a dose of 400mg of Compound L-1 and 200mg of Compound C-1.

EXAMPLE 12 bilayer tablet (controlled Release of Compound L-1/immediate Release of Compound C-1)

Compound L-1(200mg) layer				Compound C-1(200mg) layer
								Composition (I)	％w/w	mg/tablet	g/1000 tablets	Composition (I)	％w/w	mg/tablet	g/1000 tablets
Compound (I)aL-1	36.36	200.00	200.00	Compound C-1	40.00	200.00	200.00
								dHydroxypropyl methylcellulose	45.00	247.50	247.50	b，cLactose	40.75	203.75	203.75
bLactose	17.14	94.25	94.25	Sodium lauryl sulfate	3.00	15.00	15.00

Magnesium stearate	1.50	8.25	8.25	cPolyvinylpyrrolidone (PVP)	2.50	12.50	12.50
								-	-	-	-	eCroscarmellose sodium	3.00	15.00	15.00
-	-	-	-	cMicrocrystalline cellulose	10.00	50.00	50.00
								-	-	-	-	Magnesium stearate	0.75	3.75	3.75
Total amount of	100.00	550.00	550.00	Total amount of	100.00	500.00	500.00

a. Compound L-1 can be used in the form of the free acid or salt; if a salt is used, the total weight of the tablet is increased or the amount of filler (e.g., lactose) is adjusted to accommodate the weight of the counter ion.

b. Lactose may be replaced by other water-soluble excipients, such as mannitol.

c. The polyvinylpyrrolidone may be replaced by other suitable binders, such as hydroxypropyl cellulose or hydroxypropylmethyl cellulose, and the amount of microcrystalline cellulose and lactose may be varied to adjust the drug release profile.

d. The amount of hydroxypropyl methylcellulose and lactose in the L-1 layer can be varied to adjust the release profile.

e. Croscarmellose sodium may be replaced by other disintegrants such as sodium starch glycolate or cross-linked polyvinylpyrrolidone.

The components of both layers were granulated separately.

For LTB₄All the components except magnesium stearate in the compound L-1 layer were granulated by a dry granulation process or a wet granulation process. The granules are dried (if wet granulated), milled and mixed with magnesium stearate to give the final mixture.

For layer C-1, all ingredients except partially crosslinked hydroxymethyl cellulose sodium and magnesium stearate are mixed together and granulated using a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled and mixed with part or all of the croscarmellose sodium, followed by final mixing with magnesium stearate.

The two final blends are then compressed into bilayer tablets of appropriate size to obtain the desired dosage.

For a dose of 200mg of Compound L-1 and 200mg of Compound C-1, bilayer tablets having a total weight of 1050mg (comprising 550mg of Compound L-1 particles and 500mg of Compound C-1 particles) were prepared.

For a dose of 100mg of Compound L-1 and 100mg of Compound C-1, bilayer tablets were prepared having a total weight of 525mg (comprising 275mg of Compound L-1 particles and 250mg of Compound C-1 particles).

EXAMPLE 13 bilayer tablet (controlled Release of Compound L-1/immediate Release of Compound C-1)

Compound L-1(200mg) layer				Compound C-1(100mg) layer
								Composition (I)	％w/w	mg/medicineSheet	g/1000 tablets	Composition (I)	％w/w	mg/tablet	g/1000 tablets
Compound (I)aL-1	36.36	200.00	200.00	Compound C-1	25.00	100.00	100.00
								dHydroxypropyl methylcellulose	45.00	247.50	247.50	b，cLactose	53.25	213.00	213.00
bLactose	17.14	94.25	94.25	Sodium lauryl sulfate	3.00	12.00	12.00
								Magnesium stearate	1.50	8.25	8.25	cPolyvinylpyrrolidone (PVP)	2.50	10.00	10.00
-	-	-	-	eCroscarmellose sodium	3.00	12.00	12.00
								-	-	-	-	cMicrocrystalline cellulose	12.50	50.00	50.00
-	-	-	-	Magnesium stearate	0.75	3.00	3.00
								Total amount of	100.00	550.00	550.00	Total amount of	100.00	400.00	400.00

a. Compound L-1 can be used in the form of the free acid or salt; if a salt is used, the total weight of the tablet is increased or the amount of filler (e.g., lactose) is adjusted to accommodate the weight of the counter ion.

b. Lactose may be replaced by other water-soluble excipients, such as mannitol.

c. The polyvinylpyrrolidone may be replaced by other suitable binders, such as hydroxypropyl cellulose or hydroxypropylmethyl cellulose, and the amount of microcrystalline cellulose and lactose may be varied to adjust the drug release profile.

d. The amount of hydroxypropyl methylcellulose and lactose in the compound L-1 layer can be varied to adjust the release profile.

e. Croscarmellose sodium may be replaced by other disintegrants such as sodium starch glycolate or cross-linked polyvinylpyrrolidone.

The components of both layers were granulated separately.

For the compound L-1 layer, all the ingredients except magnesium stearate were granulated by a dry granulation process or a wet granulation process. The granules are dried (if wet granulated), milled and mixed with magnesium stearate to give the final mixture.

For compound C-1 layer, all ingredients except partially crosslinked hydroxymethyl cellulose sodium and magnesium stearate were mixed together and granulated using either a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled and mixed with part or all of the croscarmellose sodium, followed by final mixing with magnesium stearate.

The two final blends are then compressed into bilayer tablets of appropriate size to obtain the desired dosage.

For a dose of 200mg of Compound L-1 and 100mg of Compound C-1, bilayer tablets were prepared with a total weight of 950mg (comprising 550mg of Compound L-1 granules and 400mg of Compound C-1 granules).

Two tablets were taken to give a dose of 400mg of Compound L-1 and 200mg of Compound C-1.

EXAMPLE 14 multiparticulates (as sachet)

Compound L-1(200mg) beads (microspheres)				Compound C-1(200mg) granules
								Composition (I)	％w/w	mg/unit dose	g/1000 unit dose	Composition (I)	％w/w	mg/unit dose	g/1000 unit dose
Compound (I)aL-1	40.00	200.00	200.00	Compound C-1	62.50	200.00	200.00
								bGlyceryl behenate	57.00	285.00	285.00	c，dLactose	19.00	60.80	60.80
bPolyoxyethylene polyoxypropylene copolymer	3.00	15.00	15.00	Sodium lauryl sulfate	3.00	9.60	9.60

-	-	-	-	dPolyvinylpyrrolidone (PVP)	2.50	8.00	8.00
								-	-	-	-	eCroscarmellose sodium	3.00	9.60	9.60
-	-	-	-	dMicrocrystalline cellulose	10.00	32.00	32.00
								Total amount of	100.00	500.00	500.00	Total amount of	100.0	320.00	320.00

a. Compound L-1 can be used in the form of the free acid or salt; if a salt is used, the total weight of the beads per dose is increased to accommodate the weight of the counter ion.

b. The polyoxyethylene polyoxypropylene copolymer may be replaced by polyglycolized glycerides (polyglycolyzedgerides) and/or the amount of glyceryl behenate may be varied to modulate the drug release rate.

c. Lactose may be replaced by other water-soluble excipients, such as mannitol.

d. The polyvinylpyrrolidone may be replaced by other suitable binders, such as hydroxypropyl cellulose or hydroxypropylmethyl cellulose, and the amount of microcrystalline cellulose and lactose may be varied to adjust the drug release profile.

e. Croscarmellose sodium may be replaced by other disintegrants such as sodium starch glycolate or cross-linked polyvinylpyrrolidone.

For compound L-1 multiparticulates, all ingredients were mixed together and then multiparticulates (or microspheres) were prepared using a melt spray freezing process. These multiparticulates can then be used "as is" or, if desired, coated with a sustained release polymer (e.g., a mixture of ethylcellulose, cellulose acetate and cellulose acetate phthalate) or an enteric coating polymer (e.g., hydroxypropylmethyl cellulose phthalate or methacrylic acid copolymer) to modify the release profile.

For compound C-1 granules, all ingredients were mixed together and granulated using a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled to the desired particle size.

Then, compound L-1 microspheres (500mg plus weight of coating polymer) and compound C-1 particles (320mg) were mixed together and 820mg (plus other weights to account for the weight of the polymer coating) of this mixture was prepared as sachets to give a dose of 200mg of compound L-1 and 200mg of compound C-1. For lower doses of 100mg of Compound L-1 and 100mg of Compound C-1, 410mg (plus additional weight to account for the weight of the polymer coating) of this mixture was used.

As an alternative to sachets, for lower doses, the mixture may be filled in capsules and taken in capsule form while capsules of acceptable size may contain the mixture.

EXAMPLE 15 multiparticulates (as sachet)

Compound L-1(200mg) beads (microspheres)				Compound C-1(100mg) granules
								Composition (I)	％w/w	mg/unit dose	g/1000 unit dose	Composition (I)	％w/w	mg/unit dose	g/1000 unit dose
Compound (I)aL-1	40.00	200.00	200.00	Compound C-1	40.00	100.00	100.00
								bGlyceryl behenate	57.00	285.00	285.00	c，dLactose	40.50	101.25	101.25
bPolyoxyethylene polyoxypropylene copolymer	3.00	15.00	15.00	Sodium lauryl sulfate	3.00	7.50	7.50
								-	-	-	-	dPolyvinylpyrrolidone (PVP)	2.50	6.25	6.25
-	-	-	-	eCroscarmellose sodium	3.00	7.50	7.50
								-	-	-	-	dMicrocrystalline cellulose	11.00	27.50	27.50
Total amount of	100.00	500.00	500.00	Total amount of	100.0	250.00	250.00

a. Compound L-1 can be used in the form of the free acid or salt; if a salt is used, the total weight of the beads per dose is increased to accommodate the weight of the counter ion.

b. The polyoxyethylene polyoxypropylene copolymer may be replaced by a polyglycolized glyceride and/or the amount of glyceryl behenate may be varied to adjust the drug release rate.

c. Lactose may be replaced by other water-soluble excipients, such as mannitol.

d. The polyvinylpyrrolidone may be replaced by other suitable binders, such as hydroxypropyl cellulose or hydroxypropylmethyl cellulose, and the amount of microcrystalline cellulose and lactose may be varied to adjust the drug release profile.

e. Croscarmellose sodium may be replaced by other disintegrants such as sodium starch glycolate or cross-linked polyvinylpyrrolidone.

For compound L-1 multiparticulates, all ingredients were mixed together and then multiparticulates (or microspheres) were prepared using a melt spray freezing process. These multiparticulates can then be used "as is" or, if desired, coated with a sustained release polymer (e.g., a mixture of ethylcellulose, cellulose acetate and cellulose acetate phthalate) or an enteric coating polymer (e.g., hydroxypropylmethyl cellulose phthalate or methacrylic acid copolymer) to modify the release profile.

For compound C-1 granules, all ingredients were mixed together and granulated using a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled to the desired particle size.

Then, compound L-1 microspheres (500mg plus weight of coating polymer) and compound C-1 particles (250mg) were mixed together and 750mg (plus other weights to account for the weight of the polymer coating) of this mixture was prepared as sachets to give a dose of 200mg of compound L-1 and 100mg of compound C-1. For higher doses of 400mg of Compound L-1 and 200mg of Compound C-1, 1500mg (plus additional weight to account for the weight of the polymer coating) of this mixture was used.

As an alternative to sachets, for lower doses, the mixture may be filled in capsules and taken in capsule form while capsules of acceptable size may contain the mixture.

EXAMPLE 16 Capsule with two drugs mixed together during granulation

Compound L-1/Compound C-1	200/200mg			200/100mg
							Composition (I)	％w/w	mg/capsule	g/1000 capsules	％w/w	mg/capsule	g/1000 capsules
Compound (I)aL-1	41.67	200.00	200.00	52.63	200.00	200.00
							Compound C-1	41.67	200.00	200.00	26.32	100.00	100.00

b，cLactose	9.16	44.00	44.00	13.55	51.50	51.50
							cSodium Lauryl Sulfate (SLS)	3.00	14.40	14.40	3.00	11.40	11.40
cPolyvinylpyrrolidone (PVP)	2.50	12.00	12.00	2.50	9.50	9.50
							dCroscarmellose sodium	1.00	4.80	4.80	1.00	3.80	3.80
Magnesium stearate	1.00	4.80	4.80	1.00	3.80	3.80
							Total amount of	100.00	480.00	480.00	100.00	380.00	380.00

a. Compound L-1 can be used in the form of the free acid or salt; if a salt is used, the total fill weight of the capsule is increased or the amount of filler (e.g., lactose) is adjusted to accommodate the weight of the counter ion.

b. Lactose may be replaced by other water-soluble excipients, for example mannitol, the amount of which is adjusted so as to completely fill the capsule.

c. The polyvinylpyrrolidone may be replaced by other suitable binders such as hydroxypropyl cellulose, and the amount of lactose and SLS may be varied to modify the drug release profile.

d. Croscarmellose sodium may be replaced by other disintegrants such as sodium starch glycolate or cross-linked polyvinylpyrrolidone.

Compound L-1, compound C-1, lactose, SLS, PVP and croscarmellose sodium (partially or completely) are mixed together and granulated by a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled and mixed with part or all of the croscarmellose sodium, followed by final mixing with magnesium stearate. The final mixture is then filled into capsules of appropriate size and fill weight to obtain the desired dosage.

Two capsules containing 200mg of Compound L-1 and 100mg of Compound C-1 were taken at a dose of 400mg of Compound C-1 and 200mg of Compound C-1.

EXAMPLE 17 Single layer tablet with two drugs granulated together

Compound L-1/Compound C-1 dose	200/200mg			200/100mg
							Composition (I)	％w/w	mg/tablet	g/1000 tablets	％w/w	mg/tablet	g/1000 tablets
Compound (I)aL-1-EDA	32.58	228.05	228.05	38.01	228.05	228.05
							Compound C-1	28.57	200.00	200.00	16.66	100.00	100.00
b，cMannitol	29.35	205.45	205.45	35.83	214.95	214.95
							Sodium Lauryl Sulfate (SLS)	3.00	21.00	21.00	3.00	18.00	18.00
cPolyvinylpyrrolidone (PVP)	2.50	17.50	17.50	2.50	15.00	15.00
							dCroscarmellose sodium	3.00	21.00	21.00	3.00	18.00	18.00
Magnesium stearate	1.00	7.00	7.00	1.00	6.00	6.00
							Total amount of	100.00	700.00	700.00	100.00	600.00	600.00

a.228.05mg of the Compound L-1-EDA (ethylenediamine salt) corresponds to 200mg of the free acid of the Compound L-1

b. Mannitol may be replaced by other water soluble excipients. Other fillers may be added to facilitate compression.

c. The polyvinylpyrrolidone can be replaced by other suitable binders and the amount of mannitol can be varied to adjust the drug release profile.

d. Croscarmellose sodium may be replaced by other disintegrants (such as sodium starch glycolate or cross-linked polyvinylpyrrolidone) or removed for controlled release of both drugs as required for tablets.

Mixing the compound L-1-EDA, the compound C-1, mannitol, SLS, PVP and croscarmellose sodium (partially or totally) together, and granulating by dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled and mixed with part or all of the croscarmellose sodium, followed by final mixing with magnesium stearate. The final blend is then compressed into tablets of appropriate size to obtain the desired dosage.

Two tablets containing 200mg of Compound L-1 and 100mg of Compound C-1 were taken at a dose of 400mg of Compound C-1 and 200mg of Compound C-1.

EXAMPLE 18 bilayer tablet (controlled Release of Compound L-1/immediate Release of Compound C-1)

Compound L-1(200mg) layer				Compound C-1(200mg) layer
								Composition (I)	％w/w	mg/tablet	g/1000 tablets	Composition (I)	％w/w	mg/tablet	g/1000 tablets
Compound (I)aL-1-EDA	38.01	228.05	228.05	Compound C-1	40.00	200.00	200.00
								dPolyethylene oxide	30.66	183.94	183.94	b，cMannitol	40.75	203.75	203.75
Mannitolb	14.33	86.00	86.00	Sodium lauryl sulfate	3.00	15.00	15.00
								Sodium chloride	14.99	89.95	89.95	cPolyvinylpyrrolidone (PVP)	2.50	12.50	12.50
Butylated hydroxytoluene	0.01	0.060	0.060	eCroscarmellose sodium	3.00	15.00	15.00
								Silicon dioxide	0.50	3.00	3.00	cMicrocrystalline cellulose	10.00	50.00	50.00
Magnesium stearate	1.50	9.00	9.00	Magnesium stearate	0.75	3.75	3.75
								Total amount of	100.00	600.00	600.00	Total amount of	100.00	500.00	500.00

a.228.05mg of the Compound L-1-EDA (ethylenediamine salt) corresponds to 200mg of the free acid of the Compound L-1

b. Mannitol may be replaced by other water-soluble excipients such as polyethylene glycol and sodium chloride. Other fillers may be added to facilitate compression.

c. The polyvinylpyrrolidone can be replaced by other suitable binders and the amounts of microcrystalline cellulose and mannitol can be varied to adjust the drug release profile.

d. The polyoxyethylene may be a mixture of more than one molecule with a molecular weight of 200000-5 million.

e. Croscarmellose sodium may be replaced by other disintegrants such as sodium starch glycolate or cross-linked polyvinylpyrrolidone.

The components of both layers were granulated separately.

For the compound L-1 layer, all ingredients except for silicon dioxide and magnesium stearate were granulated using a dry granulation process or a wet granulation process. The granules are dried (if wet granulated), milled and mixed with silicon dioxide, followed by final mixing with magnesium stearate.

For compound C-1 layer, all ingredients except partially crosslinked hydroxymethyl cellulose sodium and magnesium stearate were mixed together and granulated using either a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled and mixed with part or all of the croscarmellose sodium, followed by final mixing with magnesium stearate.

The two final blends are then compressed into bilayer tablets of appropriate size to obtain the desired dosage.

For a dose of 200mg of Compound L-1 and 200mg of Compound C-1, a bilayer tablet having a total weight of 1100mg (comprising 600mg of Compound L-1 granules and 500mg of Compound C-1 granules) was prepared.

For a dose of 100mg of Compound L-1 and 100mg of Compound C-1, bilayer tablets were prepared with a total weight of 550mg (comprising 300mg of Compound L-1 granules and 250mg of Compound C-1 granules).

EXAMPLE 19 bilayer tablet (controlled Release of Compound L-1/immediate Release of Compound C-1)

Compound L-1(200mg) layer				Compound C-1(200mg) layer
								Composition (I)	％w/w	mg/tablet	g/1000 tablets	Composition (I)	％w/w	mg/tablet	g/1000 tablets
Compound (I)aL-1-EDA	38.01	228.05	228.05	Compound C-1	25.00	100.00	100.00

dPolyethylene oxide	30.66	183.94	183.94	b，cMannitol	53.25	213.00	213.00
								Mannitolb	14.33	86.00	86.00	Sodium lauryl sulfate	3.00	12.00	12.00
Sodium chloride	14.99	89.95	89.95	cPolyvinylpyrrolidone (PVP)	2.50	10.00	10.00
								Butylated hydroxytoluene	0.01	0.06	0.06	eCroscarmellose sodium	3.00	12.00	12.00
Silicon dioxide	0.50	3.00	3.00	cMicrocrystalline cellulose	12.50	50.00	50.00
								Magnesium stearate	1.50	9.00	9.00	Magnesium stearate	0.75	3.00	3.00
Total amount of	100.00	600.00	600.00	Total amount of	100.00	400.00	400.00

a.228.05mg of the Compound L-1-EDA (ethylenediamine salt) corresponds to 200mg of the free acid of the Compound L-1

b. Mannitol may be replaced by other water-soluble excipients such as polyethylene glycol and sodium chloride. Other fillers may be added to facilitate compression.

c. The polyvinylpyrrolidone can be replaced by other suitable binders and the amounts of microcrystalline cellulose and mannitol can be varied to adjust the drug release profile.

d. The polyoxyethylene may be a mixture of more than one molecule with a molecular weight of 200000-5 million.

e. Croscarmellose sodium may be replaced by other disintegrants such as sodium starch glycolate or cross-linked polyvinylpyrrolidone.

The components of both layers were granulated separately.

For the compound L-1 layer, all ingredients except for silicon dioxide and magnesium stearate were granulated using a dry granulation process or a wet granulation process. The granules are dried (if wet granulated), milled and mixed with silicon dioxide, followed by final mixing with magnesium stearate.

For compound C-1 layer, all ingredients except partially crosslinked hydroxymethyl cellulose sodium and magnesium stearate were mixed together and granulated using either a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled and mixed with part or all of the croscarmellose sodium, followed by final mixing with magnesium stearate.

The two final blends are then compressed into bilayer tablets of appropriate size to obtain the desired dosage.

For a dose of 200mg of Compound L-1 and 100mg of Compound C-1, bilayer tablets were prepared with a total weight of 1000mg (comprising 600mg of Compound L-1 granules and 400mg of Compound C-1 granules).

Two tablets were taken so that the dose was 400mg of Compound L-1 and 200mg of Compound C-1.

EXAMPLE 20 multiparticulates (e.g. as sachets)

Compound L-1(200mg) beads (microspheres)				Compound C-1(200mg) granules
								Composition (I)	％w/w	mg/unit dose	g/1000 unit dose	Composition (I)	％w/w	mg/unit dose	g/1000 unit dose
Compound (I)aL-1-EDA	45.61	228.05	228.05	Compound C-1	62.50	200.00	200.00
								bGlyceryl behenate	51.39	256.95	256.95	c，dMannitol	19.00	60.80	60.80
bPolyoxyethylene polyoxypropylene copolymer	3.00	15.00	15.00	Sodium lauryl sulfate	3.00	9.60	9.60
								-	-	-	-	dPolyvinylpyrrolidone (PVP)	2.50	8.00	8.00
-	-	-	-	eCroscarmellose sodium	3.00	9.60	9.60
								-	-	-	-	dMicrocrystalline cellulose	10.00	32.00	32.00
Total amount of	100.00	500.00	500.00	Total amount of	100.0	320.00	320.00

a.228.05mg of compound L-1-EDA (ethylenediamine salt) corresponds to 200mg of compound L-1 free acid.

b. The polyoxyethylene polyoxypropylene copolymer may be replaced by a polyglycolized glyceride and/or the amount of glyceryl behenate may be varied to adjust the drug release rate.

c. Mannitol may be replaced by other water-soluble excipients.

d. The polyvinylpyrrolidone may be replaced by other suitable binders and the amount of microcrystalline cellulose and lactose may be varied to adjust the drug release profile.

e. Croscarmellose sodium may be replaced by other disintegrants such as sodium starch glycolate or cross-linked polyvinylpyrrolidone.

For compound L-1 multiparticulates, all ingredients were mixed together and then multiparticulates (or microspheres) were prepared using a melt spray freezing process. These multiparticulates can then be used "as is" or, if desired, coated with a sustained release polymer (e.g., a mixture of ethylcellulose, cellulose acetate and cellulose acetate phthalate) or an enteric coating polymer (e.g., hydroxypropylmethyl cellulose phthalate or methacrylic acid copolymer) to modify the release profile.

For compound C-1 granules, all ingredients were mixed together and granulated using a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled to the desired particle size.

Then, compound L-1 microspheres (500mg plus weight of coating polymer) and compound C-1 particles (320mg) were mixed together and 820mg (plus other weights to account for the weight of the polymer coating) of this mixture was prepared as sachets to give a dose of 200mg of compound L-1 and 200mg of compound C-1. For lower doses of 100mg of Compound L-1 and 100mg of Compound C-1, 410mg (plus additional weight to account for the weight of the polymer coating) of this mixture was used.

As an alternative to sachets, for lower doses, the mixture may be filled in capsules and taken in capsule form while capsules of acceptable size may contain the mixture.

EXAMPLE 21 multiparticulates (e.g. as sachets)

Compound L-1(200mg) beads (microspheres)				Compound C-1(100mg) granules
								Composition (I)	％w/w	mg/unit dose	g/1000 unit dose	Composition (I)	％w/w	mg/unit dose	g/1000 unit dose
Compound (I)aL-1-	45.61	228.05	228.05	Compound C-1	40.00	100.00	100.00

EDA
								bGlyceryl behenate	51.39	256.95	256.95	c，dLactose	40.50	101.25	101.25
bPolyoxyethylene polyoxypropylene copolymer	3.00	15.00	15.00	Sodium lauryl sulfate	3.00	7.50	7.50
								-	-	-	-	dPolyvinylpyrrolidone (PVP)	2.50	6.25	6.25
-	-	-	-	eCroscarmellose sodium	3.00	7.50	7.50
								-	-	-	-	dMicrocrystalline cellulose	11.00	27.50	27.50
Total amount of	100.00	500.00	500.00	Total amount of	100.0	250.00	250.00

a.228.05mg of compound L-1-EDA (ethylenediamine salt) corresponds to 200mg of compound L-1 free acid.

b. The polyoxyethylene polyoxypropylene copolymer may be replaced by a polyglycolized glyceride and/or the amount of glyceryl behenate may be varied to adjust the drug release rate.

c. Lactose may be replaced by other water-soluble excipients.

d. The polyvinylpyrrolidone can be replaced by other suitable binders and the amounts of microcrystalline cellulose and mannitol can be varied to adjust the drug release profile.

e. Croscarmellose sodium may be replaced by other disintegrants such as sodium starch glycolate or cross-linked polyvinylpyrrolidone.

For compound L-1 multiparticulates, all ingredients were mixed together and then multiparticulates (or microspheres) were prepared using a melt spray freezing process. These multiparticulates can then be used "as is" or, if desired, coated with a sustained release polymer (e.g., a mixture of ethylcellulose, cellulose acetate and cellulose acetate phthalate) or an enteric coating polymer (e.g., hydroxypropylmethyl cellulose phthalate or methacrylic acid copolymer) to modify the release profile.

For compound C-1 granules, all ingredients were mixed together and granulated using a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially not mixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled to the desired particle size.

Then, compound L-1 microspheres (500mg plus weight of coating polymer) and compound C-1 particles (250mg) were mixed together and 750mg (plus other weights to account for the weight of the polymer coating) of this mixture was prepared as sachets to give a dose of 200mg of compound L-1 and 100mg of compound C-1. For higher doses of 400mg of Compound L-1 and 200mg of Compound C-1, 1500mg (plus additional weight to account for the weight of the polymer coating) of this mixture was used.

As an alternative to sachets, for lower doses, the mixture may be filled in capsules and taken in capsule form while capsules of acceptable size may contain the mixture.

EXAMPLE 22 Capsule with two drugs mixed together in granules

Compound L-1/Compound C-1 dose	200/200mg			200/100mg
							Composition (I)	％w/w	mg/capsule	g/1000 capsules	％w/w	mg/capsule	g/1000 capsules
Compound (I)aL-1-EDA	45.61	228.05	228.05	60.01	228.05	228.05
							Compound C-1	40.00	200.00	200.00	26.32	100.00	100.00
b，cMannitol	6.89	34.45	34.45	6.17	23.45	23.45
							cSodium Lauryl Sulfate (SLS)	3.00	15.00	15.00	3.00	11.40	11.40
cPolyvinylpyrrolidone (PVP)	2.50	12.50	12.50	2.50	9.50	9.50
							dCroscarmellose sodium	1.00	5.00	5.00	1.00	3.80	3.80
Magnesium stearate	1.00	5.00	5.00	1.00	3.80	3.80
							Total amount of	100.00	5.00	5.00	100.00	380.00	380.00

a.228.05mg of compound L-1-EDA (ethylenediamine salt) corresponds to 200mg of compound L-1 free acid.

b. Mannitol may be replaced by other water-soluble fillers, the amount of which is adjusted to completely fill the capsule.

c. The polyvinylpyrrolidone may be replaced by other suitable binders and the amount of mannitol and SLS may be varied to modify the drug release profile.

d. Croscarmellose sodium may be replaced by other disintegrants such as sodium starch glycolate or cross-linked polyvinylpyrrolidone.

Compound L-1-EDA, compound C-1, mannitol, SLS, PVP and croscarmellose sodium (partially or totally) are mixed together and granulated using a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled and mixed with part or all of the croscarmellose sodium, followed by final mixing with magnesium stearate. The final mixture is then filled into capsules of appropriate size and fill weight to obtain the desired dosage.

Two capsules containing 200mg of Compound L-1 and 100mg of Compound C-1 were taken at a dose of 400mg of Compound C-1 and 200mg of Compound C-1.

The examples herein may also be carried out by substituting the reactants and/or operating conditions described in the preceding examples with those described generally or specifically.

Thus, as the invention is described, it is obvious that the same may be varied in some ways. Such variations are not to be regarded as a departure from the spirit and scope of the invention, and all such modifications and equivalents as would be obvious to one skilled in the art are intended to be included within the scope of the following claims.

EXAMPLE 23 double layer tablet

Compound L-1(200mg) layer				Compound C-1(200mg) layer
								Composition (I)	％w/w	mg/tablet	g/1000 tablets	Composition (I)	％w/w	mg/tablet	g/1000 tablets
Compound (I)aL-1	40.00	200.00	200.00	Compound C-1	40.00	200.00	200.00
								b，cLactose	42.00	210.00	210.00	b, c lactose	40.75	203.75	203.75
Sodium Lauryl Sulfate (SLS)	1.00	5.00	5.00	Sodium Lauryl Sulfate (SLS)	3.00	15.00	15.00

cPolyvinylpyrrolidone (PVP)	2.50	12.50	12.50	cPolyvinylpyrrolidone (PVP)	2.50	12.50	12.50
								dCroscarmellose sodium	3.00	15.00	15.00	dCroscarmellose sodium	3.00	15.00	15.00
cMicrocrystalline cellulose	10.00	50.00	50.00	cMicrocrystalline cellulose	10.00	50.00	50.00
								Magnesium stearate	1.50	7.50	7.50	Magnesium stearate	0.75	3.75	3.75
Total amount of	100.00	500.00	500.00	Total amount of	100.00	500.00	500.00

a. Compound L-1 can be used in the form of the free acid or salt; if a salt is used, the total weight of the tablet is increased or the amount of filler (e.g., lactose) is adjusted to accommodate the weight of the counter ion.

b. Lactose may be replaced by other water-soluble excipients, such as mannitol.

c. The polyvinylpyrrolidone can be replaced by other suitable binders such as hydroxypropyl cellulose or hydroxypropyl methyl cellulose, and the amounts of binder, microcrystalline cellulose and lactose can be varied to adjust the drug release characteristics and compression characteristics of the tablet.

d. Croscarmellose sodium may be replaced by other disintegrants (such as sodium starch glycolate or cross-linked polyvinylpyrrolidone), and the amount may be adjusted to adjust the disintegration time.

The components of both layers were granulated separately.

For compound L-1 layer, all ingredients except croscarmellose sodium and magnesium stearate were mixed together and granulated using a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled and mixed with part or all of the croscarmellose sodium, followed by final mixing with the remaining magnesium stearate.

For compound C-1 layer, all ingredients except partially crosslinked hydroxymethyl cellulose sodium and magnesium stearate were mixed together and granulated using either a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled and mixed with part or all of the croscarmellose sodium, followed by final mixing with the remaining magnesium stearate.

The two final blends are then compressed into bilayer tablets of appropriate size to obtain the desired dosage.

For a dose of 200mg of Compound L-1 and 200mg of Compound C-1, bilayer tablets were prepared with a total weight of 1000mg (comprising 500mg of Compound L-1 granules and 500mg of Compound C-1 granules).

For a dose of 100mg of Compound L-1 and 100mg of Compound C-1, bilayer tablets were prepared with a total weight of 500mg (comprising 250mg of Compound L-1 granules and 250mg of Compound C-1 granules).

EXAMPLE 25 bilayer tablets

Compound L-1(200mg) layer				Compound C-1(100mg) layer
								Composition (I)	％w/w	mg/tablet	g/1000 tablets	Composition (I)	％w/w	mg/tablet	g/1000 tablets
Compound (I)aL-1	40.00	200.00	200.00	Compound C-1	25.00	100.00	100.00
								b，cLactose	42.00	210.00	210.00	b，cLactose	53.25	213.00	213.00
Sodium Lauryl Sulfate (SLS)	1.00	5.00	5.00	Sodium Lauryl Sulfate (SLS)	3.00	12.00	12.00
								cPolyvinylpyrrolidone (PVP)	2.50	12.50	12.50	cPolyvinylpyrrolidone (PVP)	2.50	10.00	10.00
dCroscarmellose sodium	3.00	15.00	15.00	dCroscarmellose sodium	3.00	12.00	12.00
								cMicrocrystalline cellulose	10.00	50.00	50.00	cMicrocrystalline cellulose	12.50	50.00	50.00
Magnesium stearate	1.50	7.50	7.50	Magnesium stearate	0.75	3.00	3.00
								Total amount of	100.00	500.00	500.00	Total amount of	100.00	400.00	400.00

a. Compound L-1 can be used in the form of the free acid or salt; if a salt is used, the total weight of the tablet is increased or the amount of filler (e.g., lactose) is adjusted to accommodate the weight of the counter ion.

b. Lactose may be replaced by other water-soluble excipients, such as mannitol.

c. The polyvinylpyrrolidone can be replaced by other suitable binders such as hydroxypropyl cellulose or hydroxypropyl methyl cellulose, and the amounts of binder, microcrystalline cellulose and lactose can be varied to adjust the drug release characteristics and compression characteristics of the tablet.

d. Croscarmellose sodium may be replaced by other disintegrants (such as sodium starch glycolate or cross-linked polyvinylpyrrolidone), and the amount may be adjusted to adjust the disintegration time.

The components of both layers were granulated separately.

For compound L-1 layer, all ingredients except croscarmellose sodium and magnesium stearate were mixed together and granulated using a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled and mixed with part or all of the croscarmellose sodium, followed by final mixing with the remaining magnesium stearate.

For compound C-1 layer, all ingredients except partially crosslinked hydroxymethyl cellulose sodium and magnesium stearate were mixed together and granulated using either a dry granulation or wet granulation process. For wet granulation, some or all of the SLS and/or PVP is initially unmixed with the drug and added as a solution during the granulation step. If desired, the granules are comminuted and dried (for wet granulation). The dried granules are milled and mixed with part or all of the croscarmellose sodium, followed by final mixing with the remaining magnesium stearate.

The two final blends are then compressed into bilayer tablets of appropriate size to obtain the desired dosage.

For a dose of 200mg of Compound L-1 and 100mg of Compound C-1, bilayer tablets were prepared with a total weight of 900mg (comprising 500mg of Compound L-1 granules and 400mg of Compound C-1 granules).

Two tablets were taken so that the dose was 400mg of Compound L-1 and 200mg of Compound C-1.

Example 25 murine Air Pouch model

Female Balb/c mice for 8-12 weeks were injected subcutaneously with approximately 5 ml of air using a 10 ml syringe with a 27 gauge needle and a 0.2 micron Acrodisc filter. Animal quilt CO₂/O₂The mixture was paralyzed and then injected subcutaneously with air in the scapular region of the mice. The animal's balloon was re-inflated with about 2-3 ml of air every 2-3 days, as above. The fifth morning at 6 am dosing was initiated with each compound including vehicle (0.5% methylcellulose + 0.025% Tween 80) or COX-2 inhibitor (Compound C-1, 15mpk, BID), or LTB₄Receptor antagonists (Compound L-2, 150mpk, BID), or COX-2 inhibitors and LTB₄Combinations of receptor antagonists (Compound C-1, 30mpk + Compound L-2, 150mpk, BID). Compounds were administered orally and quantitatively in a volume of 0.2 ml per dose. On day 7, animals were again CO-fed₂/O₂Paralyzing, and the air cell was injected with a stimulus (1% zymosan sigma Z-4250) prepared in saline solution (0.9% saline, Baxter #2f7122) using a 27 gauge pillow and a 3 ml syringe. Two hours after stimulation, mice were euthanized and the tape was lavaged with a 1ml volume of Dmen/F12Gibco # 21041-025. Then, the cells were placed in test tubes in ice to maintain the gastric lavage fluid of each isolated mouse. Cells were centrifuged at 1600rpm for 10 minutes. The cells were then resuspended in 1ml of the same medium used for lavage of the cells. Total cells/mouse were calculated using a coulter counter. The results of this study are listed in table 7, corresponding to figure 1.

TABLE 7

Group of	Total cells/mouse	Standard deviation of	SEM
				Control	51800000	179000	8000
Vehicle + Zymosan	43000000	1410	812
				30mpk COX-2 inhibitors	40600000	16200	7230
300mpk LTB4RA	38900000	22000	11000
				COX-2 inhibitor + LTB4 RA	24300000	15000	7510

Standard error of SEM-mean

Claims (41)

1. A therapeutic composition comprising at least one COX-2 selective inhibitor or prodrug thereof and at least one LTB₄A receptor antagonist, wherein the LTB₄The receptor antagonist comprises one or more compounds selected from the group consisting of 2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl]-4- (trifluoromethyl) benzoic acid, pharmaceutically acceptable salts thereof and mixtures thereof.

2. The therapeutic composition of claim 1 wherein the COX-2 selective inhibitor comprises celecoxib.

3. The therapeutic composition of claim 1, wherein the LTB₄The receptor antagonist comprises 2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl]-4- (trifluoromethyl) benzoic acid.

4. The therapeutic composition of claim 3 wherein the COX-2 selective inhibitor comprises celecoxib.

5. The therapeutic composition of claim 1, wherein the LTB₄The receptor antagonist comprises 2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl]A pharmaceutically acceptable salt of (E) -4- (trifluoromethyl) benzoic acid.

6. The therapeutic composition of claim 1, wherein the LTB₄The receptor antagonist comprises 2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl]Mono (ethylenediamine) salt of-4- (trifluoromethyl) benzoic acid.

7. The therapeutic composition of claim 6 wherein the COX-2 selective inhibitor comprises celecoxib.

8. The composition of claim 1 or 2, further comprising a pharmaceutically acceptable excipient.

9. The composition of claim 1 or 2, wherein the composition is a solid dosage form.

10. The composition of claim 9, wherein the solid dosage form is an oral dosage form.

11. The composition of claim 10, wherein the oral dosage form is selected from the group consisting of tablets, capsules, suppositories, pills, gel caps, and granular compositions.

12. The composition of claim 11, wherein the oral dosage form is a capsule.

13. The composition of claim 12, wherein the capsule is an extended release capsule dosage form.

14. The composition of claim 11, wherein the oral dosage form is a tablet dosage form.

15. The composition of claim 14, wherein the tablet dosage form is selected from the group consisting of a multilayer tablet dosage form, a sustained release tablet dosage form, a core shell tablet dosage form, an osmotic tablet dosage form, and a side-by-side tablet dosage form.

16. The composition of claim 15, wherein said tablet dosage form comprises a multilayer tablet dosage form.

17. The therapeutic composition of claim 15 wherein the tablet dosage form comprises a parallel tablet dosage form.

18. The composition of claim 15, wherein said tablet dosage form comprises a sustained release tablet dosage form.

19. The composition of claim 15, wherein the tablet dosage form comprises a core shell tablet dosage form.

20. The composition of claim 1 or 2, wherein the COX-2 selective inhibitor or prodrug thereof and LTB₄The receptor antagonist is present in an intimate mixture.

21. The composition of claim 1 or 2, wherein the composition is in an aqueous dosage form.

22. The composition of claim 21, wherein the aqueous dosage form is a syrup.

23. The composition of claim 21, wherein the aqueous dosage form is suitable for parenteral administration.

24. The composition of claim 1 or 2, wherein the composition is an inhalable dosage form.

25. The composition of claim 1 or 2, wherein the composition is a semi-solid dosage form.

26. The composition of claim 25, wherein the semi-solid dosage form is suitable for topical application.

27. The composition of claim 1 or 2, wherein the composition is a suspension.

28. A method for treating, preventing or inhibiting inflammation, an inflammation-related disorder, a pain-related disorder or pain in a subject in need thereof, said method comprising administering to said subject a composition comprising a COX-2 selective anti-inflammatory compound and an LTB₄Compositions of receptor antagonist compounds wherein the LTB₄The receptor antagonist compound comprises one or more compounds selected from the group consisting of 2- [ (3S, 4R) -3, 4-dihydro-4-hydroxy-3- (phenylmethyl) -2H-1-benzopyran-7-yl]-4- (trifluoromethyl) benzoic acid, salts thereof and mixtures thereof.

29. The method of claim 28, wherein said COX-2 selective inhibitor comprises celecoxib.

30. The method of any one of claims 28 or 29, wherein the target object is an animal.

31. The method of claim 30, wherein the animal is a human.

32. The method of claim 31, for treating, preventing or inhibiting a disorder associated with inflammation.

33. The method of claim 31, for treating, preventing or inhibiting inflammation.

34. The method of claim 31 for treating, preventing or inhibiting pain.

35. The method of claim 31 for treating, preventing or inhibiting a pain-associated disorder.

36. The method of claim 31, wherein the inflammation-related disorder is arthritis.

37. The method of claim 36, wherein the arthritis is osteoarthritis.

38. The method of claim 37, wherein the arthritis is rheumatoid arthritis.

39. The method of claim 31 for the prevention or treatment of any one or more of the disorders selected from the group consisting of: connective tissue and joint disorders, neoplasia disorders, cardiovascular disorders, ear disorders, eye disorders, respiratory disorders, gastrointestinal disorders, disorders related to angiogenesis, immunological disorders, allergic disorders, nutritional disorders, infectious diseases and disorders, endocrine disorders, metabolic disorders, neurological and neurodegenerative disorders, psychiatric disorders, liver and bile disorders, musculoskeletal disorders, genitourinary disorders, gynecological and obstetrical disorders, injury and trauma disorders, surgical disorders, dental and oral disorders, sexual disorders, skin disorders, blood disorders and toxic disorders.

40. The method of claim 28, wherein the LTB is administered in an amount that is sequential in nature₄A receptor antagonist and an amount of a COX-2 selective inhibitor.

41. The method of claim 28, wherein the amount of LTB is administered in a substantially simultaneous manner₄A receptor antagonist and an amount of a COX-2 selective inhibitor.