CA2247121A1 - Mercapto derivatives as inhibitors of cyclooxygenases - Google Patents
Mercapto derivatives as inhibitors of cyclooxygenases Download PDFInfo
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- CA2247121A1 CA2247121A1 CA 2247121 CA2247121A CA2247121A1 CA 2247121 A1 CA2247121 A1 CA 2247121A1 CA 2247121 CA2247121 CA 2247121 CA 2247121 A CA2247121 A CA 2247121A CA 2247121 A1 CA2247121 A1 CA 2247121A1
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Abstract
This invention is directed to a pharmacologically acceptable composition for inhibiting cyclooxygenase in a mammal, which includes a mercapto derivative and a pharmaceutically acceptable carrier. The invention also concerns a method of inhibiting cyclooxygenase and treating various conditions where there is an advantage in inhibiting prostaglandins biosynthesis. The method includes the step of administering to a mammal a mercapto derivative in pure form or in a pharmaceutically acceptable carrier.
Description
W O 97/32575 PCTrUS97/03279 MERCAPTO DERIVATIVES AS
INHIBITORS OF CYCLOOXYGENASES
Related Aoplication This application is a continuation in part of Application Seriai No. 08/410,312 entitled "~VIERCAPTO DERIVATIVES AS
INHIBITORS OF NITRIC OXIDE SYNTHASE" filed on March 24, ~ 995, which is incorporated herein in its entirety by reference.
Backqround of the Invention The present invention relates to the use of mercapto derivatives as inhibitors of cyclooxygenases (COX).
Prostaglandins (PG's~ are synthesized from arachidonic acid by a family of enzymes termed cyclooxygenases (COX). One isoform (COX-1) is constitutively present in a variety of tissues and releases PG's in low amounts. The continuous release of PG from COX-1 serves physiological purposes. For instance, prostacyclin, a ,. vasodilatory and anti-aggregatory prostaglandin, reduces the adhesion of platelets to the endothelial surface.
CA 02247l2l ~
~ ~ . . ~ . ... ..
The inducible isoform of COX ~(CCIX-2~.is~ expre~sèd- ln .- ~.
response to immunological stimuli in multiple cell types inciuding macrophages, vascular smooth muscle cells and epithelial cells, and produces large amounts of PG's, which can result in tissue injury.
There is substantial evidence that COX-2 plays an important role in the pathogensis of a variety of inflammatory conditions. In animal models of endotoxic shock, endotoxin causes induction of COX-2. In addition, it is now thought that excess PG
production may be involved in a number of other inflammatory conditions, including arthritis and uicerative colitis.
Various inhibitors of COX have been proposed for therapeutic use. For example, non-steroidal anti-inflammatory drugs (acetylsalicylic acid, ibuprofen, etc.) are inhibitors of COX. Examples of United States patents directed to COX inhibition include 5,155,110;
5,360,925; 5,399,970; 5,409,944; 5,474,995; and 5,475,021.
Although the COX inhibitors discussed above have therapeutic use, it is important to identify additional compounds which inhibit COX. It also is desirable to identify additional compounds which may have combined actions, i.e., inhibiting the activity of COX, as well as of ~ ,~
other pro-inflammatory enzymes.
British Journal o~ Pharmacology, 117, Feb. 1996, pp. 619-632, G.J. Southan et al., discloses preparations ~or in~usion comprising mercaptoethylguanidine (MEG). The preparations are used to inhibit nitric oxide synthase.
EP-A-558468, discloses mercapto derivatives like MEG and their use in pharmaceu~ical compositions for inhibiting nitric oxide ~ormation.
~ AMENDED SHEEr 2a ~- '-WO-A-9630007 (published 3/10/96) is directed to a pharmaceutical composition including a mercapto derivative ~or inhibition o~ nitric oxidase.
US 5360925 relates to a combination AB, A
representing a cyclooxygenase inhibitor and B representing an L-~orm o~ an arginine analogue. The combinations are said to have a dual biological activity in that they similarly inhibit both NO synthase and cyclooxygenase.
Summarv of the Invention This invention is directed to a pharmacologically acceptable composition for inhibiting COX in a mammal. The composition includes a mercapto derivative and a pharmaceutically ~ ~.
~' ~~' AMENDED SI~ET
W O 97132575 PCTrUS97/03279 acceptable carrier, with the mercapto derivative present in the composition in an effective amount to inhibit COX in the mammal.
In this regard, the above-identified patent application relates to the use of the same mercapto compounds in the treatment of conditions associated with the overproduction of nitric oxide, a potent cytotoxic free radical. Thus, the mercapto compounds as COX
inhibitors described in the present application have a distinct advantage over other classes of COX inhibitors, in that they also inhibit another class of infla",n,atory enzymes (nitric oxide synthases).
The invention also is directed to a method of inhibiting COX in a mammal, which includes the step of administering to the mammal a mercapto derivative in a pure form or in a pharmaceutically acceptable carrier.
The mercapto compound or derivative of the composition and method is defined by a formula selected from the group consisting of:
R, - Y - Z - X - C
and C - X - Z - S - S - Z - X - C
Il I .
or a salt thereof, wherein W 097/32575 PCTrUS97/03279 R1 is H, alkyl, alkenyl, phenyl, alkylene, alkenylene, or phenylalkylene or a substituted derivative thereof;
When Rl is alkylene or alkenylene, R1 optionally may be joined to either of the amidino Ns, to Z or to X of the above formula containing R1 to form a 5-, 6- or 7- membered heterocyclic ring, with the proviso that~, when R1 is attached to Z, Z is alkylene or alkenylene or a substituted derivative thereof, and, when Rl is attached to X, X is either CPs5 or N;
R2, R3, R ' 2 and R 3 are independently H, lower alkyi, alkenyl, alkylene, slkenylene, amino, phenyl or phenylalkylene, or a substituted derivative thereof;
When R2 or R ' 2 iS alkylene or alkenylene, R2 or R ' 2 optionally may be joined to the imino N residing on the adjacent amidino C to form a 5- or 6- membered heterocyclic ring;
Z and Z' are independently alkylene, alkenylene, cycioalkylene or cycloalkenylene, or a substituted derivative thereof;
When R2, R3, R 2 or R'3 is alkylene or alkenylene, R2, R3, R 2 or R'3 optionaily may be joined to the adjacent Z or Z' to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of 0 or S, with the proviso that said Z or Z' so joined is an alkylene or alkenylene, said heterocyclic ring optionally being substituted with a lower alkyl, alkoxy, halo, hydroxy or amino;
X is N, NR4, 0, CR5 or CR4R5; ~.
X is N, NR 4, 0, CR'5 or CR'4R 5;
W 097/32575 PCT~US97/03279 Y is S;
R4 and R'4 are independently H, alkyl, alkylene, alkenylene, thioalkylene or thioesteralkylene;
R5 and R ' 5 are independently H, alkyl, alkylene, alkenylene, thioalkylene, thioesteralkylene, amino or carboxyl; and When R4 or R'4 is alkylene, alkenylene, thioalkylene, or thioesteralkylene, R4 or R '4 optionally may be joined to R2, R3, R '2 or R'3 to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of 0 or S, with the proviso that said R2, R3, R'2 or R'3 so joined is alkylene, alkenylene, amino, phenyl, phenylalkylene, or a substituted derivative thereof wherein the substituted derivative is lower alkyl or halo.
Brief Desc.i,.tion of the Drawinqs Fig. 1 is a graph of the effect of mercaptoethylguanidine (MEG) on 6-keto prostaglandin F1 alpha production by (A) non-stimulated macrophages in the presence of arachidonic acid, and (B) immunostimulated J774 macrophages (stimulated with LPS and interferon-gamma for 6h).
Fig. 2 is a graph of the effect of mercaptoethylguanidine (MEG) on thromboxane B2 production by (A) non-stimulated macrophages in the presence of arachidonic acid, and (B) immunostimulated J774 macrophages (stimulated with LPS and t interferon-gamma for 6h).
_ WO 97132575 PCT~US97/03279 Fig. 3 is a graph of the effect of mercaptoethylguanidine (MEG), on 6-keto prostaglandin F1 aipha production by (A) purified COX-1, and (B) purified COX-2 in a cell-free system (N=3-6).
Detaiied Descril~tion of the Invention This invention is directed to a pharmacologically acceptable composition for inhibiting COX in a mammal. The composition includes a mercapto derivative and a pharmaceutically acceptable carrier, with the mercapto derivative present in the composition in an effective amount to inhibit COX in the mammal. The invention also is directed to a method of inhibiting COX in a mammal, which includes the step of administering to the mammal a mercapto derivative in pure form or in a pharmaceutically acceptable carrier.
Suitable mercapto derivatives for use in the composition or method may be made according to the methods of synthesis taught in the following articles which are incorporated herein in their entirety by reference:
(1) Joseph X. Khym et al., "lon Exchange Studies of Transguanylation Reactions. I. Rearrangement of S,2-Aminoethylisothiourea to 2-Mercaptoethylguanidine and 2-Aminothiazoline", Journal of the American Chemical Society, Vol. 79, pp. 5663-5666, November 5, 1957;
INHIBITORS OF CYCLOOXYGENASES
Related Aoplication This application is a continuation in part of Application Seriai No. 08/410,312 entitled "~VIERCAPTO DERIVATIVES AS
INHIBITORS OF NITRIC OXIDE SYNTHASE" filed on March 24, ~ 995, which is incorporated herein in its entirety by reference.
Backqround of the Invention The present invention relates to the use of mercapto derivatives as inhibitors of cyclooxygenases (COX).
Prostaglandins (PG's~ are synthesized from arachidonic acid by a family of enzymes termed cyclooxygenases (COX). One isoform (COX-1) is constitutively present in a variety of tissues and releases PG's in low amounts. The continuous release of PG from COX-1 serves physiological purposes. For instance, prostacyclin, a ,. vasodilatory and anti-aggregatory prostaglandin, reduces the adhesion of platelets to the endothelial surface.
CA 02247l2l ~
~ ~ . . ~ . ... ..
The inducible isoform of COX ~(CCIX-2~.is~ expre~sèd- ln .- ~.
response to immunological stimuli in multiple cell types inciuding macrophages, vascular smooth muscle cells and epithelial cells, and produces large amounts of PG's, which can result in tissue injury.
There is substantial evidence that COX-2 plays an important role in the pathogensis of a variety of inflammatory conditions. In animal models of endotoxic shock, endotoxin causes induction of COX-2. In addition, it is now thought that excess PG
production may be involved in a number of other inflammatory conditions, including arthritis and uicerative colitis.
Various inhibitors of COX have been proposed for therapeutic use. For example, non-steroidal anti-inflammatory drugs (acetylsalicylic acid, ibuprofen, etc.) are inhibitors of COX. Examples of United States patents directed to COX inhibition include 5,155,110;
5,360,925; 5,399,970; 5,409,944; 5,474,995; and 5,475,021.
Although the COX inhibitors discussed above have therapeutic use, it is important to identify additional compounds which inhibit COX. It also is desirable to identify additional compounds which may have combined actions, i.e., inhibiting the activity of COX, as well as of ~ ,~
other pro-inflammatory enzymes.
British Journal o~ Pharmacology, 117, Feb. 1996, pp. 619-632, G.J. Southan et al., discloses preparations ~or in~usion comprising mercaptoethylguanidine (MEG). The preparations are used to inhibit nitric oxide synthase.
EP-A-558468, discloses mercapto derivatives like MEG and their use in pharmaceu~ical compositions for inhibiting nitric oxide ~ormation.
~ AMENDED SHEEr 2a ~- '-WO-A-9630007 (published 3/10/96) is directed to a pharmaceutical composition including a mercapto derivative ~or inhibition o~ nitric oxidase.
US 5360925 relates to a combination AB, A
representing a cyclooxygenase inhibitor and B representing an L-~orm o~ an arginine analogue. The combinations are said to have a dual biological activity in that they similarly inhibit both NO synthase and cyclooxygenase.
Summarv of the Invention This invention is directed to a pharmacologically acceptable composition for inhibiting COX in a mammal. The composition includes a mercapto derivative and a pharmaceutically ~ ~.
~' ~~' AMENDED SI~ET
W O 97132575 PCTrUS97/03279 acceptable carrier, with the mercapto derivative present in the composition in an effective amount to inhibit COX in the mammal.
In this regard, the above-identified patent application relates to the use of the same mercapto compounds in the treatment of conditions associated with the overproduction of nitric oxide, a potent cytotoxic free radical. Thus, the mercapto compounds as COX
inhibitors described in the present application have a distinct advantage over other classes of COX inhibitors, in that they also inhibit another class of infla",n,atory enzymes (nitric oxide synthases).
The invention also is directed to a method of inhibiting COX in a mammal, which includes the step of administering to the mammal a mercapto derivative in a pure form or in a pharmaceutically acceptable carrier.
The mercapto compound or derivative of the composition and method is defined by a formula selected from the group consisting of:
R, - Y - Z - X - C
and C - X - Z - S - S - Z - X - C
Il I .
or a salt thereof, wherein W 097/32575 PCTrUS97/03279 R1 is H, alkyl, alkenyl, phenyl, alkylene, alkenylene, or phenylalkylene or a substituted derivative thereof;
When Rl is alkylene or alkenylene, R1 optionally may be joined to either of the amidino Ns, to Z or to X of the above formula containing R1 to form a 5-, 6- or 7- membered heterocyclic ring, with the proviso that~, when R1 is attached to Z, Z is alkylene or alkenylene or a substituted derivative thereof, and, when Rl is attached to X, X is either CPs5 or N;
R2, R3, R ' 2 and R 3 are independently H, lower alkyi, alkenyl, alkylene, slkenylene, amino, phenyl or phenylalkylene, or a substituted derivative thereof;
When R2 or R ' 2 iS alkylene or alkenylene, R2 or R ' 2 optionally may be joined to the imino N residing on the adjacent amidino C to form a 5- or 6- membered heterocyclic ring;
Z and Z' are independently alkylene, alkenylene, cycioalkylene or cycloalkenylene, or a substituted derivative thereof;
When R2, R3, R 2 or R'3 is alkylene or alkenylene, R2, R3, R 2 or R'3 optionaily may be joined to the adjacent Z or Z' to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of 0 or S, with the proviso that said Z or Z' so joined is an alkylene or alkenylene, said heterocyclic ring optionally being substituted with a lower alkyl, alkoxy, halo, hydroxy or amino;
X is N, NR4, 0, CR5 or CR4R5; ~.
X is N, NR 4, 0, CR'5 or CR'4R 5;
W 097/32575 PCT~US97/03279 Y is S;
R4 and R'4 are independently H, alkyl, alkylene, alkenylene, thioalkylene or thioesteralkylene;
R5 and R ' 5 are independently H, alkyl, alkylene, alkenylene, thioalkylene, thioesteralkylene, amino or carboxyl; and When R4 or R'4 is alkylene, alkenylene, thioalkylene, or thioesteralkylene, R4 or R '4 optionally may be joined to R2, R3, R '2 or R'3 to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of 0 or S, with the proviso that said R2, R3, R'2 or R'3 so joined is alkylene, alkenylene, amino, phenyl, phenylalkylene, or a substituted derivative thereof wherein the substituted derivative is lower alkyl or halo.
Brief Desc.i,.tion of the Drawinqs Fig. 1 is a graph of the effect of mercaptoethylguanidine (MEG) on 6-keto prostaglandin F1 alpha production by (A) non-stimulated macrophages in the presence of arachidonic acid, and (B) immunostimulated J774 macrophages (stimulated with LPS and interferon-gamma for 6h).
Fig. 2 is a graph of the effect of mercaptoethylguanidine (MEG) on thromboxane B2 production by (A) non-stimulated macrophages in the presence of arachidonic acid, and (B) immunostimulated J774 macrophages (stimulated with LPS and t interferon-gamma for 6h).
_ WO 97132575 PCT~US97/03279 Fig. 3 is a graph of the effect of mercaptoethylguanidine (MEG), on 6-keto prostaglandin F1 aipha production by (A) purified COX-1, and (B) purified COX-2 in a cell-free system (N=3-6).
Detaiied Descril~tion of the Invention This invention is directed to a pharmacologically acceptable composition for inhibiting COX in a mammal. The composition includes a mercapto derivative and a pharmaceutically acceptable carrier, with the mercapto derivative present in the composition in an effective amount to inhibit COX in the mammal. The invention also is directed to a method of inhibiting COX in a mammal, which includes the step of administering to the mammal a mercapto derivative in pure form or in a pharmaceutically acceptable carrier.
Suitable mercapto derivatives for use in the composition or method may be made according to the methods of synthesis taught in the following articles which are incorporated herein in their entirety by reference:
(1) Joseph X. Khym et al., "lon Exchange Studies of Transguanylation Reactions. I. Rearrangement of S,2-Aminoethylisothiourea to 2-Mercaptoethylguanidine and 2-Aminothiazoline", Journal of the American Chemical Society, Vol. 79, pp. 5663-5666, November 5, 1957;
(2) David G. Doherty, et al., "Synthesis of Aminoalkylisothiuronium Salts and their Conversion to Mercaptoalkylguanidines and Thiazolines", Journal of the American Chemlcal Society, Vol. 79, pp. 5667-5671, November 5, 1957;
(3) Joseph X. Khym, et al., "lon Exchange Studies of Transguanylation Reactions. Il. Rearrangement of 3-Aminopropylisothiourea and N-Substituted Aminoethyl- and Aminopropylisothioureas to Mercaptoalkylguanidines and 2-W 097/32~75 PCTnUS97/03279 -7-Aminothiazolines or Penthiazolines", Journa/of the American Chemical Society, Vol. 80, pp. 3342-3349, July 5, 1958;
[~ David G. Doherty et al. "Synthesis of D- and L-2-Aminobutylisothiourea Dihydrobromide Isomers and their Conversion to Guanidothiols, Disulfides, and Thiazolines", Journa/ of Organic Chemistry, Voi. 28, pp. 1339-1342, 1g63.
(5) Shih-Hsi Chu et al., "Potential ~ntiradiation Agents.
Il. Selenium Analogs of 2-Arrlinoethylisothiouronium Hydrobromide and Related Compounds", Journal of the American Chemical Society, Vol. 27, pp. 2899-2901, August, 1962.
(63 Tohru Hino et al., "Radiation-protective Agents. I.
Studies on N-Alkylated-2-(2-aminoethyl~thiopseudoureas and 1,1 -(Dithioethylene~diguanidines", Chemical & Pharmaceutical Bul/etin, Vol. 14, No. 11, pp. 1193-1201, November, 1966.
Suitable mercapto derivatives also may be made according to the examples provided at the end of this detailed description of the invention .
The mercapto derivative of the composition and method is defined by a formula selected from the group consisting of:
INl - R3 Rl - Y - Z - X - C
and NH - R 2 INl - R3 C - X - Z - S - S - Z - X - C
N - R'3 NH - R2 or a salt thereof, wherein Rl is H, alkyl, alkenyl, phenyl, alkylene, alkenylene, or phenylalkylene or a substituted derivative thereof;
W O 97/32575 PCT~US97/03~79 When R1 is alkylene or alkenylene, R1 optionally may be joined to either of the amidino Ns, to Z or to X of the above formula containing R1 to form a 5-, 6- or 7- membered heterocyclic ring, with the proviso that, when Rl is attached to Z, Z is alkylene or alkenylene or a substituted derivative thereof, and, when R1 is attached to X, X is either CR5 or N;
R2, R3, R'2 and R'3 are independently H, lower alkyl, alkenyl, alkylene, alkenylene, amino, phenyl or phenylalkylene, or a substituted derivative thereof;
When R2 or R'2 is alkylene or alkenylene, R2 or R'2 optionally may be joined to the imino 1~1 residing on the adjacent amidino C to form a 5- or 6- membered heterocyclic ring;
Z and Z' are independently alkylene, alkenylene, cycloalkylene or cycloalkenylene, or a substituted derivative thereof;
When R2, R3, R'2 or R'3 is alkylene or alkenylene, R2, R3, R'2 or R'3 optionally may be joined to the adjacent Z or Z' to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of 0 or S, with the proviso that Z is an alkylene or alkenylene, said heterocyclic ring optionally being substituted with a lower alkyl, alkoxy, halo, hydroxy or amino;
X is N, NR4, 0, CR5 or CR4R5;
X' is N, NR'4, 0, CR'5 or CR'4R'5;
Y is S;
W O 97/32575 PCT~US97/03279 g R4 and R'4 are independently H, alkyl, alkylene, aikenylene, thioalkylene or thioesteralkylene;
R5 and R'5 are independently H, alkyl, alkyiene, alkenylene, thioalkylene, thioesteralkyiene, amino or carboxyl; and When R4 or R'4 is alkylene, alkenylene, thioalkylene, or thioesteralkylene, R4or R'4 optionally may be joined to R2, R3, R'2 or R'3 to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of 0 or S, with the proviso that R2, R3, R '2 or R ' 3 is alkylene, alkenylene, amino, phenyl, phenylalkylene, or a substituted derivative thereof wherein the substituted derivative is lower alkyl or halo.
As used herein, the term "salt" refers to any addition salt derived from any pharmaceutically acceptable organic or inorganic acid.
Examples of suitable acids include hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene p sulfonic, tartaric, acetic, citric, methanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic and benzenesulphonic acids. Additionally, as used herein, any alkyl or alkylene may be straight chain, branched or cyclic, and "halo" includes bromine, chlorine, fluorine and iodine.
As mentioned above, Rl is H, alkyl, alkenyl, phenyl, alkylene, alkenylene or phenylalkylene, or a substituted derivative thereof. If desired, this R1 derivative may be substituted with one or more alkoxy, halo, hydroxy, amino or nitro groups. Additionally, as W O 9~/3257~ PCTrUS97/03279 noted above, R2, R3, R'2 and R'3 are independently H, lower aikyl, alkenyl, alkylene, alkenylene, amino, phenyl or phenylalkylene, or a substituted derivative thereof. If desired, the R2, R3, R'2 and R'3 derivative may be substituted with a lower alkyl or halo.
If the R4, R5, R'4 or R'5 substituent is thioalkylene, the thioalkylene preferably has a formula [-(CH2)n-SH] where n is independently 1 to 4. If R4, R5, R'4 or R'5 is thioesteralkylene, the thioesteralkylene preferably has the formula ~-(CH2)n-S-R6] where R6 is independently a lower alkyl and n is independently 1 to 4.
The Z and Z' substituents of the mercapto derivative are independently alkylene, alkenylene, cycloalkylene or cycloalkenyiene, or a substituted derivative thereof. When such a substituted derivative is employed, the substituent may include an alkoxy, halo, hydroxy, amino or nitro group.
~ A preferred subgroup of the mercapto derivative includes mercapto derivatives where: R1 is H or lower alkyl; R2 is H; R3 is 11; R'2 is H; R'3 is H; X is NR4; X' is NR'4; R4 and R'4 are independently H, methyl or ethyl; and Z and Z' are independently alkylene. A few nonlimiting examples include mercaptoethylguanidine, mercaptopropylguanidine, S-methyl-mercaptoethylguanidine, S-methyl-mercaptopropylguanidine, and guanidinoethyldisulfide. Another preferred subgroup of mercapto derivatives is formed wherein: R1 is H;
R2 is H; R3 is H; R'2 is H; R'3 is H; X is NR4; X' is NR'4; R4 is H; R'4 is H; and Z and Z' are independently a Cl 6 alkylene. Nonlimiting W O 97/3257~ PCT~US97/03279 examples include mercaptoethylguanidine, mercaptopropylguanidine, and guanidinoethyldisulfide.
The mercapto derivative, in pure form or in a pharmaceutically acceptable carrier, will find benefit in treating conditions and disorders where there is an advantage in inhibiting the cyclooxygenase enzymes. For example, the mercapto derivative may be used to treat a circulatory shock including its various aspects such as vascular and myocardial dysfunction, metabolic failure including the inhibition of mitochondrial enzymes and cytochrome P450-mediated drug metabolism, and multiple organ dysfunction syndrome including adult respiratory distress syndrome. Circulatory shock may be a result of gram-negative and gram positive sepsis, trauma, hemorrhage, burn injury, anaphylaxis, cytokineimmunotherapy, liverfailure, kidneyfailure or systemic inflammatory response syndrome. Mercapto derivatives also may be beneficial for patients receiving therapy with cytokines such as TNF, IL-1 and IL-2 or therapy with cytokine-inducing agents, or as an adjuvant to short term immunosuppression in transplant therapy. In addition, mercapto derivatives may be useful to inhibit PG
synthesis in patients suffering from inflammatory conditions in which COX-2 activity contributes to the pathophysiology of the condition, such as arthritis, inflammatory bowel disease, and myocarditis, for example.
There is also evidence that COX-2 may be involved in the pathophysiology of autoimmune and/or inflammatory conditions such CA 02247l2l l998-08-2l W O 97/3257~ PCTrUS97/032n as arthritis, rheumatoid arthritis and systemic lupus erythematosus (SLE) and in insulin-dependent diabetes mellitus, and, therefore, mercapto derivatives may prove helpful in treating these conditions.
Furthermore, it is now clear that there are a number of additional inflammatory and noninflammatory diseases that are associated with COX-2 induction. Examples of such physiological disorders include: inflammatory bowel diseases such as ileitis and Crohn's disease; inflammatory lung disorders such as asthma and chronic obstructive airway disease; inflammatory disorders of the eye including corneal dystrophy, trachoma, onchocerciasis, uveitis, sympathetic ophthalmitis and endophthalmitis; chronic inflammatory disorders of the gum including periodontitis; chronic inflammatory disorders of theioints including arthritis and osteoarthritis, tuberculosis, leprosy, glomerulonephritis sarcoid, and nephrosis; disorders of the skin 1 5 including sclerodermatitis, psoriasis and eczema; infiammatory diseases of the central nervous system, including chronic demyelinatin~ diseases such as multiple sclerosis, dementia including AlDS-related neurodegeneration and Alzheimer's disease, encephalomyelitis and viral or autoimmune encephalitis; autoimmune diseases including immune-complex vasculitis, systemic lupus and erythematodes; and disease of the heart including ischemic heart disease and cardiomyopathy.
Additional diseases which may benefit from the use of mercapto derivatives include adrenal insufficiency; hypercholesterolemia;
atherosclerosis; bone disease associated with increased bone CA 02247121 l99X-08-21 W O 97132575 PCTrUS97/03279 resorption, e.g., osteoporosis, pre-eclampsia, eclampsia, uremic complications; chronic liver failure, noninflammatory diseases of the central nervous system (CNS) including stroke and cerebral ischemia;
and various forms of cancer.
Pharmaceutical formulations of the mercapto derivative may include those suitable for oral, rectal, nasal, topical (including buccal and sub-lingual~, vaginal or parenteral (including intramuscular, sub-cutaneous and intravenous) administration, or for administration by inhalation or insufflation. The formulations may, where appropriate, be conveniently presented in discrete dosage units and may be prepared by any of the methods well known in the art of pharmacy. All such pharmacy methods include the steps of bringing into association the active compound with liquid carriers or finely divided solid carriers or both as needed and then, if necessary, shaping the product into the desired formulation.
Pharmaceutical formulations suitable for oral administration may conveniently be presented: as discrete units, such as capsules, cachets or tablets, each containing a predetermined amount of the active ingredient; as a powder or granules; or as a solution, a suspension or as an emulsion. The active ingredient may also be presented as a bolus electuary or paste, and be in a pure form, i.e., without a carrier. Tablets and capsules for oral administration may contain conventional excipients such as binding agents, fillers, lubricants, disintegrant or wetting agents. A tablet may be made by CA 02247l2l l998-08-2l W O 97/32575 PCT~US97/03279 compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredients in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diiuent, lubricating, surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may be coated according to methods well known in the art. Oral fluid preparations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations may contain conventional additives such as suspending agents, emulsifying agents, non-aqueous vehicles (which may include edible oils), or preservatives. The tablets may optionally be formulated so as to provide slow or controlled release of the active ingredient therein.
Formulations for parenteral administration include:
aqueous and non-aqueous sterile in~ection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The formulations may be presented in unit dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (Iyophilized) W 097/32575 PCTnUS97103279 condition requiring only the addition of the sterile liquid carrier, for example, saline, water-for-injection, immediately prior to use.
Alternatively, the formulations may be presented for continuous infusion. Extemporaneous injection solutions and suspensions may be prepared from steriie powders, granules and tablets of the kind previously described.
Formulations for rectal administration may be presented as a suppository with the usual carriers such as cocoa butter or polyethylene glycol. Formulations for topical administration in the mouth, for example buccally or sublingually, include lozenges, comprising the active ingredient in a flavored base such as sucrose and acacia or tragacanth, and pastilles comprising the active ingredient in a base such as gelatin and glycerin or sucrose and acacia. For intra-nasal administration the compounds of the invention may be used as a liquid spray or dispersible powder or in the form of drops. Drops may be formulated with an aqueous or non-aqueous base also comprising one or more dispersing agents, solubilizing agents or suspending agents. Liquid sprays are conveniently delivered from pressurized packs.
For administration by inhalation the compounds according to the invention are conveniently delivered from an insufflator, nebulizer pressurized packs or other convenient means of delivering an aerosol spray. Pressurized packs may comprise a suitable propellant such as dichlorodifluoromethane, trichiorofluoromethane, W O 97/32575 PCT~US97/03279 dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount.
Alternatively, for administration by inhalation or insufflation, the compounds according to the invention may take the form of a dry powder composition, for example a powder mix of the compound and a suitable powder base such as lactose or starch. The powder composition may be presented in unit dosage form, in for example, capsules, cartridges, gelatin or blister packs from which the powder may be administered with the aid of an inhalator or insufflator.
When desired the above descrit~ed formulations, adapted to give sustained release of the active ingredient, may be employed.
The pharmaceutical compositions according to the invention may also contain other active ingredients such as antimicrobial agents, 1 ~ immunosuppressants or preservatives.
The compounds of the invention may also be used in combination with other therapeutic agents, for example, anti-inflammatory agents, particularly nitric oxide synthase inhibitors, superoxide or peroxynitrite scavengers, vasodilator prostaglandins including prostacyclin and prostaglandin El, cancer chemotherapeutic agents including cisplatin, N0 donors or N0 inhalation therapy, or PAF
- receptor antagonists.
It should be understood that in addition to the ingredients particularly mentioned above, the formulations of this invention may '' CA 02247121 1998-08-21 ~ - 1 7- ; . . . .
include other agents conventional in the art having regard to the type of formulation in question, for example, those suitable for oral administration may include flavoring agents.
Preferred unit dosage formulations are those containing an effective dose, as recited below, or an appropriate fraction thereof, of the active ingredient.
For each of the aforementioned conditions, the mercapto derivative may be administered orally or via injection at a dose of from 0.1 to 250 mg/kg per day. The dose range for aduit humans is generally from 5 mg to 17.5 g/day, preferably !~ mg to 10 g/day and most preferably 100 mg to 3 g/day. Tablets or other forms of presentation provided in discrete units may conveniently contain an amount which is effective at such dosage or as a multiple of the same, for instance, units containing 5 mg to 500 mg, usually around 100 mg 1 5 to 500 mg.
The pharmaceutical composition preferably is ad,~,i.,i~Lered orally or by injection (intravenous or subcutaneous), and the precise amount administered to a patient will be the responsibility of the at~endant physician. However, the dose employed will depend upon a number of factors, including the age and sex of the patient, the precise disorder being treated, and its severity. Also the route of administration may vary depending upon the condition and its severity.
The following Examples are provided by way of illustration .
A~ENDED Slt~ET
CA 02247121 l99X-08-21 W O 97/32575 PCT~US97/03279 This example (Fig. 1 ) illustrates the effect of mercaptoethyguanidine on arachidonic acid or immunostimulatjon-induced 6-keto prostaglandin F1 alpha formation in J774.2 macrophages. J774 macrophage cell lines were obtained from the American Type Culture Collection (ATCC) and were grown using standard methods in Dulbecco's Modified Eagle Medium IDMEM) supplemented with 10% fetal bovine serum, glutamine, penicillin ( 10,000 U/l~ and streptomycin ( 1 0,000 U/l) . Cells were grown in 96-well plates for measure of the production of prostaglandin metabolites and cell viability. All the experiments were carried out without fetal calf serum in order to avoid interference with radioimmunoassay. Concentration of 6-keto prostaglandin 1 alpha, the stable metabolite product of prostacyclin in the culture medium, was determined by radioimmunoassay. Supernatant or reaction samples were diluted 1:5 in a buffer containing 0.1% polyvinylpyrolidine, 0.9%
NaCI, ~;0 mM Tris base, 1.7 mM MgS04 and 0.16 mM CaCI2 (pH 7.4) before radioimmunoassay. The stable metabolite of prostacyclin, 6-keto-PGF1 alpha, was determined by radioimmunoassay as described (Wise WC, Cook JA, Haluskha PV. "Arachidonic Acid Metabolism in Endotoxin Tolerance", Adv. Shock, Vol. 10, pp. 131-142, 1983) Measurernent of cyclooxygenase 1 activity in J774 cells Cells were plated in 96-well plates at 90-100% confluence (200,uM-3mM). Inhibitors were given as a 30-minute pretreatment.
W 097/32575 PCT~US97/03279 -19-Cells were then stimulated with arachidonic acid (16 ~M) in order to activate the constitutive cyclooxygenase (COX-1). Cells were then incubated for a further period of 30 minutes and the supernatant was collected for the measurement of arachidonic acid metabolism evaiuation by radioimmunoassay.
Measurement of cyclooxygenase ~ activity in J774 cells ~:ells were place in 96-well plates at 90-100% confluence (200 ,uL final volume) and pretreated for 30 minutes with MEG (1 ,uM-3mM). Inhibitors were given as a 30-minute pretreatment. Cells were then stimulated with endotoxin of E.coli(LPS, 1 0,ug/mL) and interferon y (IFN, 150 Uml) in order to induce the expression of the inducible cyclooxygenase (COX-2). Cells were then incubated for a further period of 6 hours and the supernatant was collected for the measurement of arachidonic acid metabolism evaluation by 1 5 radioimmunoassay.
Mitochondrial respiration, an indicator of cell viability, was assessed by the mitochondrial-dependent reduction of MTT [3- (4,5 -dimethylthiazol-2-yl) - 2,5 - diphen~lLella~olium bromide] to formazan.
Cells in 96-well piates were incubated (37~ C) with MTT (0.2 mg/ml for 60 minutes). Culture medium was removed by aspiration and the r.ells solubilized in dimethylsulfoxide (DMS0) ( 100 ,ul) . The extent of reduction of MTT to formazan within cells was qua,-LiL~led by measurement of OD550 using a microplate reader. The calibration curve for the reduction of MTT to formazan was prepared in DMS0.
W097/32575 PCTrUS97/03279 Formazan production by cells was expressed as a percentage of the vaiues obtained from untreated cells.
Celis stimulated with arachidonic acid (panel a~ or endotoxin (panel b) released 6-keto prostagJandin 1 alpha into the 5culture medium. This was dose-dependently inhibited by the mercapto derivatives, mercaptoethylguanidine (MEG). The inhibition of PG
production was not due to cell killing, as these agents in their effective doses did not decrease cellular viability (not shown~. Similar to MEG, other related mercapto derivatives exhibited inhibition of COX-2 activity 10to a variable degree (Table 1).
Half-maximal inhibitory pot~ ;e s of sel~cled ~elc~,to derivatives on COX-2 activity in immunost;mulated J744 ...acr~ z~a~
Compound EC50 (~M3 S-methyl-MEG 40 N-methyl-MEG 55 MPG 5~
EXAMPLE Z
This example (Fig. 2~ illustrates the effect of mercaptoethylguanidine on arachidonic acid or immunostimulation-induced thromboxane B2 formation in J774.2 macrophages. J774 macropha~e cell lines were cultured and treated as described in CA 02247l2l l998-08-2l W O 97/32~75 -21- PCTAUS97/03279 example 1. Concentration of thromboxane B2, the stable metabolite of thromboxane A2 was determined by radioimmunoassay.
Supernatant or reaction samples were diluted 1:5 in a buffer containing 0.1% polyvinylpyrolidine, 0.9% NaCI, 50mM Tris base, 1.7 mM MgSo4 and 0 16 mM CaCI2 (pH 7.4) before radioimmunoassay. Thromboxane B2, the stable metabolite o~ thromboxane A2, was determined by radioimmunoassay as described (Wise WC, Cook JA, Haluskha PV, "Arachidonic Acid l\lletaboiism in Endotoxin Tolerance", Adv. Shock, Vol. 10, pp. 131-142, 1983).
Cells stimulated with arachidonic acid (A) or endotoxin (B) released thromboxane A2 into the culture medium. This was dose-dependently inhibited by the mercapto derivatives, mercaptoethylguanidine ~MEG). The inhibition of PG production was not due to cell killing, as these agents in their effective doses did not decrease cellular viability (not shown~.
This example (Fig. 3) illustrates the effect of mercaptoethylguanidine on 6-keto prostaglandin F1 alpha formation by purified COX-1 1 (A) and COX-2 ~B) isoenzymes. In a test tube containing 0.1 M Tris-HCL buffer (pH 8.0, final volume 2 mL) with 1 mM EDTA, o.2 mM phenol and 1 ,uM hemin, 10 units of COX-1 of COX-2 were allowed to react with 100 ,~IM arachidonic acid for 2 minutes at 37~ C in the presence or absence of MEG (1 ~M-3mM).
The reaction was quenched by addition of 50,uL stannous chloride W 097J32575 PCT~US97/03279 solution (100 mg/ml in 1 M HCI). the reaction was allowed to proceed for an additional 10 minutes and it was stopped thereafter by addition of 5 mL of a buffer contafnin~ 0.1% polyvinylpyrolidine, 0.9% NaCI, 50 mM Tris base, 1.7 mM MgSo4 and 0.16 mM CaCi2 (pH 7.4).
Concentration of 6-keto prostaglandin 1 alpha, the stable metabolite product of prostacyclin in the culture medium, was determined by radioimmunoassay. Supernatant or reaction samples were diluted 1:5 in a buffer containing 0.1% polyvinylpyrolidine, 0.9%
NaCI, 50 mM Tris base, 1.7 mM MgSo4 and Q.16 mM CaCI2 (pH 7.4) before radioimmunoassay. The stable metabolite of prostacyclin, 6-keto-PGF1 a, was determined by radioimmunoassay as described (Wise WC, Cook JA, Haluskha PV, "Arachidonic Acid Metabolism in Endotoxin Tolerancen, Adv. Shock, Vol. 10, pp. 131-142, 1983).
There was a significant production of 6-keto prostaglandin 1 alpha into the reaction mixture. This was dose-dependently inhibited by the mercapto derivatives, mercaptoethylguanidine (MEG).
This Example illustrates a method for synthesizing mercaptoethylguanidine suiphate. Mercaptoethylarnine hydrochloride (2g) was dissoived in methanol (5 ml~ and cooled in a salt/ice bath. A
cold solution of potassium hydroxide (0.99 g) ;n methanol (10 ml) was added and the mixture stirred. After 1 hour, the solution was filtered and S-methylisothiourea t29) was added to 12 ml of the filtrate. The solution was stirred at room temperature (18~C) for 16 hours under 23~
~,, , , ~ . ....... ..
nitrogen. The solution then was filtered and ether was added to precipitate the crude product which was then recrystallized from an ether/ethanol mixture.
Ah~EhfDED SHE~T
[~ David G. Doherty et al. "Synthesis of D- and L-2-Aminobutylisothiourea Dihydrobromide Isomers and their Conversion to Guanidothiols, Disulfides, and Thiazolines", Journa/ of Organic Chemistry, Voi. 28, pp. 1339-1342, 1g63.
(5) Shih-Hsi Chu et al., "Potential ~ntiradiation Agents.
Il. Selenium Analogs of 2-Arrlinoethylisothiouronium Hydrobromide and Related Compounds", Journal of the American Chemical Society, Vol. 27, pp. 2899-2901, August, 1962.
(63 Tohru Hino et al., "Radiation-protective Agents. I.
Studies on N-Alkylated-2-(2-aminoethyl~thiopseudoureas and 1,1 -(Dithioethylene~diguanidines", Chemical & Pharmaceutical Bul/etin, Vol. 14, No. 11, pp. 1193-1201, November, 1966.
Suitable mercapto derivatives also may be made according to the examples provided at the end of this detailed description of the invention .
The mercapto derivative of the composition and method is defined by a formula selected from the group consisting of:
INl - R3 Rl - Y - Z - X - C
and NH - R 2 INl - R3 C - X - Z - S - S - Z - X - C
N - R'3 NH - R2 or a salt thereof, wherein Rl is H, alkyl, alkenyl, phenyl, alkylene, alkenylene, or phenylalkylene or a substituted derivative thereof;
W O 97/32575 PCT~US97/03~79 When R1 is alkylene or alkenylene, R1 optionally may be joined to either of the amidino Ns, to Z or to X of the above formula containing R1 to form a 5-, 6- or 7- membered heterocyclic ring, with the proviso that, when Rl is attached to Z, Z is alkylene or alkenylene or a substituted derivative thereof, and, when R1 is attached to X, X is either CR5 or N;
R2, R3, R'2 and R'3 are independently H, lower alkyl, alkenyl, alkylene, alkenylene, amino, phenyl or phenylalkylene, or a substituted derivative thereof;
When R2 or R'2 is alkylene or alkenylene, R2 or R'2 optionally may be joined to the imino 1~1 residing on the adjacent amidino C to form a 5- or 6- membered heterocyclic ring;
Z and Z' are independently alkylene, alkenylene, cycloalkylene or cycloalkenylene, or a substituted derivative thereof;
When R2, R3, R'2 or R'3 is alkylene or alkenylene, R2, R3, R'2 or R'3 optionally may be joined to the adjacent Z or Z' to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of 0 or S, with the proviso that Z is an alkylene or alkenylene, said heterocyclic ring optionally being substituted with a lower alkyl, alkoxy, halo, hydroxy or amino;
X is N, NR4, 0, CR5 or CR4R5;
X' is N, NR'4, 0, CR'5 or CR'4R'5;
Y is S;
W O 97/32575 PCT~US97/03279 g R4 and R'4 are independently H, alkyl, alkylene, aikenylene, thioalkylene or thioesteralkylene;
R5 and R'5 are independently H, alkyl, alkyiene, alkenylene, thioalkylene, thioesteralkyiene, amino or carboxyl; and When R4 or R'4 is alkylene, alkenylene, thioalkylene, or thioesteralkylene, R4or R'4 optionally may be joined to R2, R3, R'2 or R'3 to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of 0 or S, with the proviso that R2, R3, R '2 or R ' 3 is alkylene, alkenylene, amino, phenyl, phenylalkylene, or a substituted derivative thereof wherein the substituted derivative is lower alkyl or halo.
As used herein, the term "salt" refers to any addition salt derived from any pharmaceutically acceptable organic or inorganic acid.
Examples of suitable acids include hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene p sulfonic, tartaric, acetic, citric, methanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic and benzenesulphonic acids. Additionally, as used herein, any alkyl or alkylene may be straight chain, branched or cyclic, and "halo" includes bromine, chlorine, fluorine and iodine.
As mentioned above, Rl is H, alkyl, alkenyl, phenyl, alkylene, alkenylene or phenylalkylene, or a substituted derivative thereof. If desired, this R1 derivative may be substituted with one or more alkoxy, halo, hydroxy, amino or nitro groups. Additionally, as W O 9~/3257~ PCTrUS97/03279 noted above, R2, R3, R'2 and R'3 are independently H, lower aikyl, alkenyl, alkylene, alkenylene, amino, phenyl or phenylalkylene, or a substituted derivative thereof. If desired, the R2, R3, R'2 and R'3 derivative may be substituted with a lower alkyl or halo.
If the R4, R5, R'4 or R'5 substituent is thioalkylene, the thioalkylene preferably has a formula [-(CH2)n-SH] where n is independently 1 to 4. If R4, R5, R'4 or R'5 is thioesteralkylene, the thioesteralkylene preferably has the formula ~-(CH2)n-S-R6] where R6 is independently a lower alkyl and n is independently 1 to 4.
The Z and Z' substituents of the mercapto derivative are independently alkylene, alkenylene, cycloalkylene or cycloalkenyiene, or a substituted derivative thereof. When such a substituted derivative is employed, the substituent may include an alkoxy, halo, hydroxy, amino or nitro group.
~ A preferred subgroup of the mercapto derivative includes mercapto derivatives where: R1 is H or lower alkyl; R2 is H; R3 is 11; R'2 is H; R'3 is H; X is NR4; X' is NR'4; R4 and R'4 are independently H, methyl or ethyl; and Z and Z' are independently alkylene. A few nonlimiting examples include mercaptoethylguanidine, mercaptopropylguanidine, S-methyl-mercaptoethylguanidine, S-methyl-mercaptopropylguanidine, and guanidinoethyldisulfide. Another preferred subgroup of mercapto derivatives is formed wherein: R1 is H;
R2 is H; R3 is H; R'2 is H; R'3 is H; X is NR4; X' is NR'4; R4 is H; R'4 is H; and Z and Z' are independently a Cl 6 alkylene. Nonlimiting W O 97/3257~ PCT~US97/03279 examples include mercaptoethylguanidine, mercaptopropylguanidine, and guanidinoethyldisulfide.
The mercapto derivative, in pure form or in a pharmaceutically acceptable carrier, will find benefit in treating conditions and disorders where there is an advantage in inhibiting the cyclooxygenase enzymes. For example, the mercapto derivative may be used to treat a circulatory shock including its various aspects such as vascular and myocardial dysfunction, metabolic failure including the inhibition of mitochondrial enzymes and cytochrome P450-mediated drug metabolism, and multiple organ dysfunction syndrome including adult respiratory distress syndrome. Circulatory shock may be a result of gram-negative and gram positive sepsis, trauma, hemorrhage, burn injury, anaphylaxis, cytokineimmunotherapy, liverfailure, kidneyfailure or systemic inflammatory response syndrome. Mercapto derivatives also may be beneficial for patients receiving therapy with cytokines such as TNF, IL-1 and IL-2 or therapy with cytokine-inducing agents, or as an adjuvant to short term immunosuppression in transplant therapy. In addition, mercapto derivatives may be useful to inhibit PG
synthesis in patients suffering from inflammatory conditions in which COX-2 activity contributes to the pathophysiology of the condition, such as arthritis, inflammatory bowel disease, and myocarditis, for example.
There is also evidence that COX-2 may be involved in the pathophysiology of autoimmune and/or inflammatory conditions such CA 02247l2l l998-08-2l W O 97/3257~ PCTrUS97/032n as arthritis, rheumatoid arthritis and systemic lupus erythematosus (SLE) and in insulin-dependent diabetes mellitus, and, therefore, mercapto derivatives may prove helpful in treating these conditions.
Furthermore, it is now clear that there are a number of additional inflammatory and noninflammatory diseases that are associated with COX-2 induction. Examples of such physiological disorders include: inflammatory bowel diseases such as ileitis and Crohn's disease; inflammatory lung disorders such as asthma and chronic obstructive airway disease; inflammatory disorders of the eye including corneal dystrophy, trachoma, onchocerciasis, uveitis, sympathetic ophthalmitis and endophthalmitis; chronic inflammatory disorders of the gum including periodontitis; chronic inflammatory disorders of theioints including arthritis and osteoarthritis, tuberculosis, leprosy, glomerulonephritis sarcoid, and nephrosis; disorders of the skin 1 5 including sclerodermatitis, psoriasis and eczema; infiammatory diseases of the central nervous system, including chronic demyelinatin~ diseases such as multiple sclerosis, dementia including AlDS-related neurodegeneration and Alzheimer's disease, encephalomyelitis and viral or autoimmune encephalitis; autoimmune diseases including immune-complex vasculitis, systemic lupus and erythematodes; and disease of the heart including ischemic heart disease and cardiomyopathy.
Additional diseases which may benefit from the use of mercapto derivatives include adrenal insufficiency; hypercholesterolemia;
atherosclerosis; bone disease associated with increased bone CA 02247121 l99X-08-21 W O 97132575 PCTrUS97/03279 resorption, e.g., osteoporosis, pre-eclampsia, eclampsia, uremic complications; chronic liver failure, noninflammatory diseases of the central nervous system (CNS) including stroke and cerebral ischemia;
and various forms of cancer.
Pharmaceutical formulations of the mercapto derivative may include those suitable for oral, rectal, nasal, topical (including buccal and sub-lingual~, vaginal or parenteral (including intramuscular, sub-cutaneous and intravenous) administration, or for administration by inhalation or insufflation. The formulations may, where appropriate, be conveniently presented in discrete dosage units and may be prepared by any of the methods well known in the art of pharmacy. All such pharmacy methods include the steps of bringing into association the active compound with liquid carriers or finely divided solid carriers or both as needed and then, if necessary, shaping the product into the desired formulation.
Pharmaceutical formulations suitable for oral administration may conveniently be presented: as discrete units, such as capsules, cachets or tablets, each containing a predetermined amount of the active ingredient; as a powder or granules; or as a solution, a suspension or as an emulsion. The active ingredient may also be presented as a bolus electuary or paste, and be in a pure form, i.e., without a carrier. Tablets and capsules for oral administration may contain conventional excipients such as binding agents, fillers, lubricants, disintegrant or wetting agents. A tablet may be made by CA 02247l2l l998-08-2l W O 97/32575 PCT~US97/03279 compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredients in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diiuent, lubricating, surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may be coated according to methods well known in the art. Oral fluid preparations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations may contain conventional additives such as suspending agents, emulsifying agents, non-aqueous vehicles (which may include edible oils), or preservatives. The tablets may optionally be formulated so as to provide slow or controlled release of the active ingredient therein.
Formulations for parenteral administration include:
aqueous and non-aqueous sterile in~ection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The formulations may be presented in unit dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (Iyophilized) W 097/32575 PCTnUS97103279 condition requiring only the addition of the sterile liquid carrier, for example, saline, water-for-injection, immediately prior to use.
Alternatively, the formulations may be presented for continuous infusion. Extemporaneous injection solutions and suspensions may be prepared from steriie powders, granules and tablets of the kind previously described.
Formulations for rectal administration may be presented as a suppository with the usual carriers such as cocoa butter or polyethylene glycol. Formulations for topical administration in the mouth, for example buccally or sublingually, include lozenges, comprising the active ingredient in a flavored base such as sucrose and acacia or tragacanth, and pastilles comprising the active ingredient in a base such as gelatin and glycerin or sucrose and acacia. For intra-nasal administration the compounds of the invention may be used as a liquid spray or dispersible powder or in the form of drops. Drops may be formulated with an aqueous or non-aqueous base also comprising one or more dispersing agents, solubilizing agents or suspending agents. Liquid sprays are conveniently delivered from pressurized packs.
For administration by inhalation the compounds according to the invention are conveniently delivered from an insufflator, nebulizer pressurized packs or other convenient means of delivering an aerosol spray. Pressurized packs may comprise a suitable propellant such as dichlorodifluoromethane, trichiorofluoromethane, W O 97/32575 PCT~US97/03279 dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount.
Alternatively, for administration by inhalation or insufflation, the compounds according to the invention may take the form of a dry powder composition, for example a powder mix of the compound and a suitable powder base such as lactose or starch. The powder composition may be presented in unit dosage form, in for example, capsules, cartridges, gelatin or blister packs from which the powder may be administered with the aid of an inhalator or insufflator.
When desired the above descrit~ed formulations, adapted to give sustained release of the active ingredient, may be employed.
The pharmaceutical compositions according to the invention may also contain other active ingredients such as antimicrobial agents, 1 ~ immunosuppressants or preservatives.
The compounds of the invention may also be used in combination with other therapeutic agents, for example, anti-inflammatory agents, particularly nitric oxide synthase inhibitors, superoxide or peroxynitrite scavengers, vasodilator prostaglandins including prostacyclin and prostaglandin El, cancer chemotherapeutic agents including cisplatin, N0 donors or N0 inhalation therapy, or PAF
- receptor antagonists.
It should be understood that in addition to the ingredients particularly mentioned above, the formulations of this invention may '' CA 02247121 1998-08-21 ~ - 1 7- ; . . . .
include other agents conventional in the art having regard to the type of formulation in question, for example, those suitable for oral administration may include flavoring agents.
Preferred unit dosage formulations are those containing an effective dose, as recited below, or an appropriate fraction thereof, of the active ingredient.
For each of the aforementioned conditions, the mercapto derivative may be administered orally or via injection at a dose of from 0.1 to 250 mg/kg per day. The dose range for aduit humans is generally from 5 mg to 17.5 g/day, preferably !~ mg to 10 g/day and most preferably 100 mg to 3 g/day. Tablets or other forms of presentation provided in discrete units may conveniently contain an amount which is effective at such dosage or as a multiple of the same, for instance, units containing 5 mg to 500 mg, usually around 100 mg 1 5 to 500 mg.
The pharmaceutical composition preferably is ad,~,i.,i~Lered orally or by injection (intravenous or subcutaneous), and the precise amount administered to a patient will be the responsibility of the at~endant physician. However, the dose employed will depend upon a number of factors, including the age and sex of the patient, the precise disorder being treated, and its severity. Also the route of administration may vary depending upon the condition and its severity.
The following Examples are provided by way of illustration .
A~ENDED Slt~ET
CA 02247121 l99X-08-21 W O 97/32575 PCT~US97/03279 This example (Fig. 1 ) illustrates the effect of mercaptoethyguanidine on arachidonic acid or immunostimulatjon-induced 6-keto prostaglandin F1 alpha formation in J774.2 macrophages. J774 macrophage cell lines were obtained from the American Type Culture Collection (ATCC) and were grown using standard methods in Dulbecco's Modified Eagle Medium IDMEM) supplemented with 10% fetal bovine serum, glutamine, penicillin ( 10,000 U/l~ and streptomycin ( 1 0,000 U/l) . Cells were grown in 96-well plates for measure of the production of prostaglandin metabolites and cell viability. All the experiments were carried out without fetal calf serum in order to avoid interference with radioimmunoassay. Concentration of 6-keto prostaglandin 1 alpha, the stable metabolite product of prostacyclin in the culture medium, was determined by radioimmunoassay. Supernatant or reaction samples were diluted 1:5 in a buffer containing 0.1% polyvinylpyrolidine, 0.9%
NaCI, ~;0 mM Tris base, 1.7 mM MgS04 and 0.16 mM CaCI2 (pH 7.4) before radioimmunoassay. The stable metabolite of prostacyclin, 6-keto-PGF1 alpha, was determined by radioimmunoassay as described (Wise WC, Cook JA, Haluskha PV. "Arachidonic Acid Metabolism in Endotoxin Tolerance", Adv. Shock, Vol. 10, pp. 131-142, 1983) Measurernent of cyclooxygenase 1 activity in J774 cells Cells were plated in 96-well plates at 90-100% confluence (200,uM-3mM). Inhibitors were given as a 30-minute pretreatment.
W 097/32575 PCT~US97/03279 -19-Cells were then stimulated with arachidonic acid (16 ~M) in order to activate the constitutive cyclooxygenase (COX-1). Cells were then incubated for a further period of 30 minutes and the supernatant was collected for the measurement of arachidonic acid metabolism evaiuation by radioimmunoassay.
Measurement of cyclooxygenase ~ activity in J774 cells ~:ells were place in 96-well plates at 90-100% confluence (200 ,uL final volume) and pretreated for 30 minutes with MEG (1 ,uM-3mM). Inhibitors were given as a 30-minute pretreatment. Cells were then stimulated with endotoxin of E.coli(LPS, 1 0,ug/mL) and interferon y (IFN, 150 Uml) in order to induce the expression of the inducible cyclooxygenase (COX-2). Cells were then incubated for a further period of 6 hours and the supernatant was collected for the measurement of arachidonic acid metabolism evaluation by 1 5 radioimmunoassay.
Mitochondrial respiration, an indicator of cell viability, was assessed by the mitochondrial-dependent reduction of MTT [3- (4,5 -dimethylthiazol-2-yl) - 2,5 - diphen~lLella~olium bromide] to formazan.
Cells in 96-well piates were incubated (37~ C) with MTT (0.2 mg/ml for 60 minutes). Culture medium was removed by aspiration and the r.ells solubilized in dimethylsulfoxide (DMS0) ( 100 ,ul) . The extent of reduction of MTT to formazan within cells was qua,-LiL~led by measurement of OD550 using a microplate reader. The calibration curve for the reduction of MTT to formazan was prepared in DMS0.
W097/32575 PCTrUS97/03279 Formazan production by cells was expressed as a percentage of the vaiues obtained from untreated cells.
Celis stimulated with arachidonic acid (panel a~ or endotoxin (panel b) released 6-keto prostagJandin 1 alpha into the 5culture medium. This was dose-dependently inhibited by the mercapto derivatives, mercaptoethylguanidine (MEG). The inhibition of PG
production was not due to cell killing, as these agents in their effective doses did not decrease cellular viability (not shown~. Similar to MEG, other related mercapto derivatives exhibited inhibition of COX-2 activity 10to a variable degree (Table 1).
Half-maximal inhibitory pot~ ;e s of sel~cled ~elc~,to derivatives on COX-2 activity in immunost;mulated J744 ...acr~ z~a~
Compound EC50 (~M3 S-methyl-MEG 40 N-methyl-MEG 55 MPG 5~
EXAMPLE Z
This example (Fig. 2~ illustrates the effect of mercaptoethylguanidine on arachidonic acid or immunostimulation-induced thromboxane B2 formation in J774.2 macrophages. J774 macropha~e cell lines were cultured and treated as described in CA 02247l2l l998-08-2l W O 97/32~75 -21- PCTAUS97/03279 example 1. Concentration of thromboxane B2, the stable metabolite of thromboxane A2 was determined by radioimmunoassay.
Supernatant or reaction samples were diluted 1:5 in a buffer containing 0.1% polyvinylpyrolidine, 0.9% NaCI, 50mM Tris base, 1.7 mM MgSo4 and 0 16 mM CaCI2 (pH 7.4) before radioimmunoassay. Thromboxane B2, the stable metabolite o~ thromboxane A2, was determined by radioimmunoassay as described (Wise WC, Cook JA, Haluskha PV, "Arachidonic Acid l\lletaboiism in Endotoxin Tolerance", Adv. Shock, Vol. 10, pp. 131-142, 1983).
Cells stimulated with arachidonic acid (A) or endotoxin (B) released thromboxane A2 into the culture medium. This was dose-dependently inhibited by the mercapto derivatives, mercaptoethylguanidine ~MEG). The inhibition of PG production was not due to cell killing, as these agents in their effective doses did not decrease cellular viability (not shown~.
This example (Fig. 3) illustrates the effect of mercaptoethylguanidine on 6-keto prostaglandin F1 alpha formation by purified COX-1 1 (A) and COX-2 ~B) isoenzymes. In a test tube containing 0.1 M Tris-HCL buffer (pH 8.0, final volume 2 mL) with 1 mM EDTA, o.2 mM phenol and 1 ,uM hemin, 10 units of COX-1 of COX-2 were allowed to react with 100 ,~IM arachidonic acid for 2 minutes at 37~ C in the presence or absence of MEG (1 ~M-3mM).
The reaction was quenched by addition of 50,uL stannous chloride W 097J32575 PCT~US97/03279 solution (100 mg/ml in 1 M HCI). the reaction was allowed to proceed for an additional 10 minutes and it was stopped thereafter by addition of 5 mL of a buffer contafnin~ 0.1% polyvinylpyrolidine, 0.9% NaCI, 50 mM Tris base, 1.7 mM MgSo4 and 0.16 mM CaCi2 (pH 7.4).
Concentration of 6-keto prostaglandin 1 alpha, the stable metabolite product of prostacyclin in the culture medium, was determined by radioimmunoassay. Supernatant or reaction samples were diluted 1:5 in a buffer containing 0.1% polyvinylpyrolidine, 0.9%
NaCI, 50 mM Tris base, 1.7 mM MgSo4 and Q.16 mM CaCI2 (pH 7.4) before radioimmunoassay. The stable metabolite of prostacyclin, 6-keto-PGF1 a, was determined by radioimmunoassay as described (Wise WC, Cook JA, Haluskha PV, "Arachidonic Acid Metabolism in Endotoxin Tolerancen, Adv. Shock, Vol. 10, pp. 131-142, 1983).
There was a significant production of 6-keto prostaglandin 1 alpha into the reaction mixture. This was dose-dependently inhibited by the mercapto derivatives, mercaptoethylguanidine (MEG).
This Example illustrates a method for synthesizing mercaptoethylguanidine suiphate. Mercaptoethylarnine hydrochloride (2g) was dissoived in methanol (5 ml~ and cooled in a salt/ice bath. A
cold solution of potassium hydroxide (0.99 g) ;n methanol (10 ml) was added and the mixture stirred. After 1 hour, the solution was filtered and S-methylisothiourea t29) was added to 12 ml of the filtrate. The solution was stirred at room temperature (18~C) for 16 hours under 23~
~,, , , ~ . ....... ..
nitrogen. The solution then was filtered and ether was added to precipitate the crude product which was then recrystallized from an ether/ethanol mixture.
Ah~EhfDED SHE~T
Claims (50)
1. Use of a mercapto derivative for the manufacture of a medicament for inhibiting cyclooxygenase in a mammal, wherein the mercapto derivative has a formula selected from the group consisting of:
and or a salt thereof, wherein R1 is H, alkyl, alkenyl, phenyl, alkylene, alkenylene, or phenylalkylene or a substituted derivative thereof;
When R1 is alkylene or alkenylene, R1 optionally may be joined to either of the amidino Ns, to Z or to X of the above formula containing R1 to form a 5-, 6- or 7- membered heterocyclic ring, with the proviso that, when R1 is attached to Z, Z is alkylene or alkenylene or a substituted derivative thereof, and, when R1 is attached to X, X is either GR5 or N;
R2, R3, R'2 and R'3 are independently H, lower alkyl, alkenyl, alkylene, alkenylene, amino, phenyl or phenylalkylene, or a substituted derivative thereof;
When R2 or R'2 is alkylene or alkenylene, R2 or R'2 optionally may be joined to the imino N residing on the adjacent amidino C to form a 5- or 6- membered heterocyclic ring;
Z and Z' are independently alkylene, alkenylene, cycloalkylene or cycloalkenylene, or a substituted derivative thereof;
When R2, R3, R'2 or R'3 is alkylene or alkenylene, R2, R3, R'2 or R'3 optionally may be joined to the adjacent Z or Z' to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of O or S, with the proviso that Z is an alkylene or alkenylene, said heterocyclic ring optionally being substituted with a lower alkyl, alkoxy, halo, hydroxy or amino;
X is N, NR4, O, CR5 or CR4R5;
X' is N, NR'4, O, CR'5 or CR'4R'5;
Y is S;
R4 and R'4 are independently H, alkyl, alkylene, alkenylene, thioalkylene or thioesteralkylene;
R5 and R'5 are independently H, alkyl, alkylene, alkenylene, thioalkylene, thioesteralkylene, amino or carboxyl;
When R4 or R'4 is alkylene, alkenylene, thioalkylene, or thioesteralkylene, R4 or R'4 optionally may be joined to R2, R3, R'2 or R'3 to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of O or S, with the proviso that R2, R3, R'2 or R'3 is independently alkylene, alkenylene, amino, phenyl, phenalkylene, or a substituted derivative thereof wherein the substituted derivative is lower alkyl or halo; and wherein the mercapto derivative is provided in a composition with a pharmaceutically acceptable carrier in an effective amount to inhibit cyclooxygenase in said mammal.
and or a salt thereof, wherein R1 is H, alkyl, alkenyl, phenyl, alkylene, alkenylene, or phenylalkylene or a substituted derivative thereof;
When R1 is alkylene or alkenylene, R1 optionally may be joined to either of the amidino Ns, to Z or to X of the above formula containing R1 to form a 5-, 6- or 7- membered heterocyclic ring, with the proviso that, when R1 is attached to Z, Z is alkylene or alkenylene or a substituted derivative thereof, and, when R1 is attached to X, X is either GR5 or N;
R2, R3, R'2 and R'3 are independently H, lower alkyl, alkenyl, alkylene, alkenylene, amino, phenyl or phenylalkylene, or a substituted derivative thereof;
When R2 or R'2 is alkylene or alkenylene, R2 or R'2 optionally may be joined to the imino N residing on the adjacent amidino C to form a 5- or 6- membered heterocyclic ring;
Z and Z' are independently alkylene, alkenylene, cycloalkylene or cycloalkenylene, or a substituted derivative thereof;
When R2, R3, R'2 or R'3 is alkylene or alkenylene, R2, R3, R'2 or R'3 optionally may be joined to the adjacent Z or Z' to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of O or S, with the proviso that Z is an alkylene or alkenylene, said heterocyclic ring optionally being substituted with a lower alkyl, alkoxy, halo, hydroxy or amino;
X is N, NR4, O, CR5 or CR4R5;
X' is N, NR'4, O, CR'5 or CR'4R'5;
Y is S;
R4 and R'4 are independently H, alkyl, alkylene, alkenylene, thioalkylene or thioesteralkylene;
R5 and R'5 are independently H, alkyl, alkylene, alkenylene, thioalkylene, thioesteralkylene, amino or carboxyl;
When R4 or R'4 is alkylene, alkenylene, thioalkylene, or thioesteralkylene, R4 or R'4 optionally may be joined to R2, R3, R'2 or R'3 to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of O or S, with the proviso that R2, R3, R'2 or R'3 is independently alkylene, alkenylene, amino, phenyl, phenalkylene, or a substituted derivative thereof wherein the substituted derivative is lower alkyl or halo; and wherein the mercapto derivative is provided in a composition with a pharmaceutically acceptable carrier in an effective amount to inhibit cyclooxygenase in said mammal.
2. Use of Claim 1 wherein said substituted derivative of R1 is selected from the group consisting of one or more of alkoxy, halo, hydroxy, amino and nitro
3 Use of Claim 1 or Claim 2 wherein said substituted derivative of R2, R3, R '2 or R '3 is independently selected from the group consisting of lower alkyl and halo.
4 Use of any preceding Claim wherein said R4, R5, R'4 or R'5 thioalkylene has a formula [-(CH2)n-SH] where n is independently 1 to 4.
5. Use of any one of Claims 1 to 3 wherein said R4, R5, R'4 or R'5 thioesteralkylene has a formula [-(CH2)n-S-R6]
where R6 is independently a lower alkyl and n is independently 1 to 4.
where R6 is independently a lower alkyl and n is independently 1 to 4.
6. Use of any preceding Claim wherein said substituted derivative of Z or Z' is independently selected from the group consisting of alkoxy, halo, hydroxy, amino and nitro
7. Use of Claim 1 wherein R1 is selected from the group consisting of H and lower alkyl, R2 is H, R3 is H, R'2 is H, R'3 is H, X is NR4, X' is NR'4, R4 and R'4 are independently selected from the group consisting of H, methyl and ethyl, and Z and Z' are independently alkylene.
8. Use of Claim 1 wherein R1 is H, R2 is H, R3 is H, R'2 is H, R'3 is H, X is NR4, X' is NR'4, R4 is H, R'4 is H and Z and Z' are independently a C1-6 alkylene.
9. Use of Claim 1 wherein said mercapto derivative is selected from the group consisting of mercaptoethylguanidine, mercaptopropylguanidine, S-methyl-mercaptoethylguanidine, S-methyl-mercaptopropylguanidine, and guanidinoethyldisulfide.
10. Use of any preceding Claim wherein said mercapto derivative is present in an amount sufficient to treat a condition where there is an advantage in inhibiting cyclooxygenases.
11. Use of Claim 10 wherein said condition is selected from the group consisting of circulatory shock, systemic inflammatory response syndrome, therapy with cytokines, therapy with cytokine-inducing agents, transplantation, transplant rejection, local inflammatory responses, systemic inflammation, autoimmune diseases, adult respiratory distress syndrome, arthritis, rheumatoid arthritis, diabetes mellitus, ileitis, ulcerative colitis, Crohn's disease, asthma, periodontitis, nephrosis, chronic demyelinating diseases of the nervous system, multiple sclerosis, AIDS-related complications, Alzheimer's disease, ischemic heartdisease, cardiomyopathy, adrenal insufficiency, hypercholesterolemia, atherosclerosis, bone diseases associated with increased bone resorption, pre-eclampsia, eclampsia, uremic complications, chronic liver failure, stroke, cerebral ischemia, and cancer.
12. Use of Claim 10 wherein said condition is selected from the group consisting of systemic inflammatory response syndrome and circulatory shocic.
13. Use of any preceding Claim wherein the composition is formulated for oral, rectal, nasal, topical, buccal, sub-lingual, vaginal, parental, intramuscular, sub-cutaneous, intravenous, inhalation or insufflation administration.
14 Use of any preceding Claim wherein the composition is formulated for oral administration, said carrier including an ingredient selected from the group consisting of a binding agent, filler, lubricant, disintegrant, wetting agent, inert diluent, surface active agent, dispersing agent, suspending agent, emulsifying agent, edible oil, flavoring agent and mixtures thereof.
15. Use of any one of Claims 1 to 13 wherein the composition is formulated for topical administration in the mouth, said carrier including an ingredient selected from the group consisting of a flavor, sucrose, acacia, tragacanth, gelatin, glycerin and mixtures thereof.
16. Use of any one of Claims 1 to 13 wherein the composition is formulated for nasal administration, said carrier including an ingredient selected from the group consisting of a dispersing agent, solubilizing agent, suspending agent and mixtures thereof.
17. Use of any one of Claims 1 to 13 wherein the composition is formulated for administration by inhalation, said carrier including a propellant.
18. Use of Claim 17 wherein said propellant is selected from the group consisting of dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide and and mixtures thereof.
19. Use of any one of Claims 1 to 13 wherein the composition is formulated for administration by inhalation or insufflation, said carrier including an ingredient selected from the group consisting of lactose, starch and mixtures thereof.
20. Use of any one of Claims 1 to 13 wherein the composition is formulated for parenteral administration, said carrier including an ingredient selected from the group consisting of an anti-oxidant, buffer, bacteriostat, suspending agent, thickening agent, saline, water and mixtures thereof.
21. Use of any one of Claims 1 to 13 wherein the composition is formulated for rectal administration, said carrier including an ingredient selected from the group consisting of cocoa butter, polyethylene glycol and mixtures thereof.
22. Use as claimed in any preceding Claim wherein the composition includes an ingredient selected from the group consisting of an antimicrobial agent, an immunosuppressant, a preservative and mixtures thereof.
23. Use as claimed in any preceding Claim wherein the composition is formulated for administration at a dose of from 5 mg to 17.5 g/day of said mercapto derivative.
24 Use as claimed in any preceding Claim wherein the composition is formulated for administration at a dose of from 5 mg to 10 g/day of said mercapto derivative.
25. Use as claimed in any preceding Claim wherein the composition is formulated for administration at a dose of from 100 mg to 3 g/day of said mercapto derivative.
26. A method for inhibiting cyclooxygenase in a mammal comprising:
administering to the mammal an effective amount of a compound to inhibit cyclooxygenase in the mammal, said compound having a formula selected from the group consisting of:
and or a salt thereof, wherein R1 is H, alkyl, alkenyl, phenyl, alkylene, alkenylene, or phenylalkylene or a substituted derivative thereof;
When R1 is alkylene or alkenylene, R1 optionally may be joined to either of the amidino Ns, to Z or to X of the above formula containing R1 to form a 5-, 6- or 7- membered heterocyclic ring, with the. proviso- that, when R1 is attached to Z, Z is alkylene or alkenylene or a substituted derivative thereof, and, when R1 is attached to X, X is either CR5 or N;
R2, R3, R'2 and R'3 are independently H, lower alkyl, alkenyl, alkylene, alkenylene, amino, phenyl or phenylalkylene, or a substituted derivative thereof, When R2 or R'2 is alkylene or alkenylene, R2 or R'2 optionally may be joined to the imino N residing on the adjacent amidino C to form a 5- or 6- membered heterocyclic ring;
Z and Z' are independently alkylene, alkenylene, cycloalkylene or cycloalkenylene, or a substituted derivative thereof;
When R2, R3, R'2 or R'3 is alkylene or alkenylene, R2, R3, R'2 or R'3 optionally may be joined to the adjacent Z or Z' to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of O or S, with the proviso that Z is an alkylene or alkenylene, said heterocyclic ring optionally being substituted with a lower alkyl, alkoxy, halo, hydroxy or amino;
X is N, NR4, O, CR5 or CR4R5;
X' is N, NR'4, O, CR'5 or CR'4R'5;
Y is S;
R4 and R'4 are independently H, alkyl, alkylene, alkenylene, thioalkylene or thioesteralkylene;
R5 and R'5 are independently H, alkyl, alkylene, alkenylene, thioalkylene, thioesteralkylene, amino or carboxyl; and When R4 or R'4 is alkylene, alkenylener thioalkylene, or thioesteralkylene, R4 or R'4 optionally may be joined to R2, R3, R'2 or R'3 to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of O or S, with the proviso that R2, R3, R'2 or R'3 is alkylene, alkenylene, amino, phenyl, phenylalkylene, or a substituted derivative thereof wherein the substituted derivative is lower alkyl or halo.
administering to the mammal an effective amount of a compound to inhibit cyclooxygenase in the mammal, said compound having a formula selected from the group consisting of:
and or a salt thereof, wherein R1 is H, alkyl, alkenyl, phenyl, alkylene, alkenylene, or phenylalkylene or a substituted derivative thereof;
When R1 is alkylene or alkenylene, R1 optionally may be joined to either of the amidino Ns, to Z or to X of the above formula containing R1 to form a 5-, 6- or 7- membered heterocyclic ring, with the. proviso- that, when R1 is attached to Z, Z is alkylene or alkenylene or a substituted derivative thereof, and, when R1 is attached to X, X is either CR5 or N;
R2, R3, R'2 and R'3 are independently H, lower alkyl, alkenyl, alkylene, alkenylene, amino, phenyl or phenylalkylene, or a substituted derivative thereof, When R2 or R'2 is alkylene or alkenylene, R2 or R'2 optionally may be joined to the imino N residing on the adjacent amidino C to form a 5- or 6- membered heterocyclic ring;
Z and Z' are independently alkylene, alkenylene, cycloalkylene or cycloalkenylene, or a substituted derivative thereof;
When R2, R3, R'2 or R'3 is alkylene or alkenylene, R2, R3, R'2 or R'3 optionally may be joined to the adjacent Z or Z' to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of O or S, with the proviso that Z is an alkylene or alkenylene, said heterocyclic ring optionally being substituted with a lower alkyl, alkoxy, halo, hydroxy or amino;
X is N, NR4, O, CR5 or CR4R5;
X' is N, NR'4, O, CR'5 or CR'4R'5;
Y is S;
R4 and R'4 are independently H, alkyl, alkylene, alkenylene, thioalkylene or thioesteralkylene;
R5 and R'5 are independently H, alkyl, alkylene, alkenylene, thioalkylene, thioesteralkylene, amino or carboxyl; and When R4 or R'4 is alkylene, alkenylener thioalkylene, or thioesteralkylene, R4 or R'4 optionally may be joined to R2, R3, R'2 or R'3 to form a 5- or 6- membered heterocyclic ring including N, C and not more than one atom of O or S, with the proviso that R2, R3, R'2 or R'3 is alkylene, alkenylene, amino, phenyl, phenylalkylene, or a substituted derivative thereof wherein the substituted derivative is lower alkyl or halo.
27. The method of claim 26 wherein said substituted derivative of R1 is selected from the group consisting of one or more of alkoxy, halo, hydroxy, amino and nitro.
28. The method of ciaim 26 wherein said substituted derivative of R2, R3, R'2 or R' 3 is independently selected from the group consisting of lower alkyl and halo.
29. The method of claim 26 wherein said R4, R5, R'4 or R' 5 thioalkylene has a formula [-(CH2)n-SH] where n is independently 1 to 4.
30. The method of claim 26 wherein said R4, R5, R'4 or R'5 thioesteralkylene has a formula [-(CH2)n-S-R6] where R6 is independently a lower alkyl and n is independently 1 to 4.
31. The method of claim 26 wherein said substituted derivative: of Z or Z' is independently selected from the group consisting of aikoxy, halo, hydroxy, amino and nitro.
32. The method of claim 26 wherein R1 is selected from the group consisting of H and lower alkyl, R2 is H, R3 is H, R'2 is H, R'3 is H, X is NR4, X' is NR'4, R4 and R'4 are independently selected from the group consisting of H, methyl and ethyl, and Z and Z' are independently alkylene.
33. The method of claim 26 wherein R1 is H, R2 is H, R3 is H, R'2 is H, R'3 is H, X is NR4, X ' is NR'4, R4 is H, R '4 is H and Z and Z' are independently a C1-6 alkylene.
34. The method of claim 26 wherein said compound is selected from the group consisting of mercaptoethylguanidine, mercaptopropylguanidine, S-methyl-mercaptoethylguanidine, S-methyl-mercaptopropylguanidine, and guanidinoethyldisulphide.
35. The method of claim 26 conducted for treating a condition where there is an advantage in inhibiting cyclooxygenase.
36. The method of claim 35 wherein said condition is selected from the group consisting of circulatory shock, systemic inflammatory response syndrome, therapy with cytokines, therapy with cytokine-inducing agents, transplantation, transplant rejection, local inflammatory responses, systemic inflammation, autoimmune diseases, adult respiratory distress syndrome, arthritis, rheumatoid arthritis, diabetes mellitus, ileitis, ulcerative colitis, Crohn's disease, asthma, periodontitis, nephrosis, chronic demyelinating diseases of the nervous system, multiple sclerosis, AIDS-related complications, Alzheimer's disease, ischemic heartdisease, cardiomyopathy, adrenal insufficiency, hypercholesterolemia, atherosclerosis, bone diseases associated with increased bone resorption, pre-eclampsia, eciampsia, uremic complications, chronic liver failure, stroke, cerebral ischemia, and cancer.
37. The method of claim 35 wherein said condition is selected from the group consisting of arthritis and gastrointestinal inflammatory disorders.
38. The method of claim 26 by administering said compound by a method selected from the group consisting of oral, rectal, nasal, topical, buccal, sub-lingual, vaginal, parenteral, intramuscular, sub-cutaneous, intravenous, inhalation and insufflation administration.
39. The method of claim 26 by orally administering said compound in a pharmacologically acceptable carrier, said carrier including an ingredient selected from the group consisting of a binding agent, filler, lubricant, disintegrant, wetting agent, inert diluent, surface active agent, dispersing agent, suspending agent, emulsifying agent, edible oil, flavoring agent and mixtures thereof.
40. The method of claim 26 by topically administering said compound in a pharmacologically acceptable carrier in the mouth, said carrier including an ingredient selected from the group consisting of a flavor, sucrose, acacia, tragacanth, gelatin, glycerin and mixtures thereof.
41. The method of claim 26 by nasally administering said compound in a pharmacologically acceptable carrier, said carrier including an ingredient selected from the group consisting of a dispersing agent, solubilizing agent, suspending agent and mixtures thereof.
42. The method of claim 26 by administering said compound in a pharmacologically acceptable carrier by inhalation, said carrier including a propellant.
43. The method of claim 42 wherein said propellant is selected from the group consisting of dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide and mixtures thereof.
44. The method of claim 26 by administering said compound in a pharmacologically acceptable carrier by inhalation or insufflation, said carrier including an ingredient selected from the group consisting of lactose, starch and mixtures thereof.
45. The method of claim 26 by administering said compound in a pharmacologically acceptable carrier parenterally, said carrier including an ingredient selected form the group consisting of an anti-oxidant, buffer, bacteriostat, suspending agent, thickening agent, saline, water and mixtures thereof.
46. The method of claim 26 by administering said compound in a pharmacologically acceptable carrier rectally, said carrier including an ingredient selected from the group consisting of cocoa butter, polyethylene glycol and mixtures thereof.
47. The method of claim 26 wherein said compound includes an ingredient selected from the group consisting of an antimicrobial agent, an immunosuppressant, a preservative and mixtures thereof.
48 The method of claim 26 wherein said compound is administered at a dose of from about 5 mg to about 17.5 g/day.
49. The method of claim 48 wherein said compound is administered at a dose of from about 5 mg to about 10 g/day.
50. The method of claim 49 wherein said compound is administered at a dose of from about 100 mg to about 3 g/day.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US61109796A | 1996-03-05 | 1996-03-05 | |
| US611,097 | 1996-03-05 | ||
| PCT/US1997/003279 WO1997032575A1 (en) | 1996-03-05 | 1997-03-03 | Mercapto derivatives as inhibitors of cyclooxygenases |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2247121A1 true CA2247121A1 (en) | 1997-09-12 |
Family
ID=29422937
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA 2247121 Abandoned CA2247121A1 (en) | 1996-03-05 | 1997-03-03 | Mercapto derivatives as inhibitors of cyclooxygenases |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA2247121A1 (en) |
-
1997
- 1997-03-03 CA CA 2247121 patent/CA2247121A1/en not_active Abandoned
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