AU4841793A - Method for antagonizing inositol 1,4,5-triphosphate - Google Patents
Method for antagonizing inositol 1,4,5-triphosphateInfo
- Publication number
- AU4841793A AU4841793A AU48417/93A AU4841793A AU4841793A AU 4841793 A AU4841793 A AU 4841793A AU 48417/93 A AU48417/93 A AU 48417/93A AU 4841793 A AU4841793 A AU 4841793A AU 4841793 A AU4841793 A AU 4841793A
- Authority
- AU
- Australia
- Prior art keywords
- inositol
- binding
- triphosphate
- calcium
- compound
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/74—Synthetic polymeric materials
- A61K31/795—Polymers containing sulfur
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
Description
METHOD FOR ANTAGONIZING INOSITOL 1,4,5-TRISPHOSPHATE
BACKGROUND OF THE INVENTION
This application relates to a series of diamino benzenedisulfonic acid oligomers that have demonstrated an affinity for the receptor sites of inositol 1,4,5- triphosphate (IP3) and are, therefore useful in diminishing the bioactivity of IP3, especially with regard to its effect on the release of intracellular calcium ions.
DESCRIPTION OF THE PRIOR ART The diamino benzenedisulfonic acid oligomers demonstrating utility as IP3 antagonists according to this invention are described in detail in the European Patent Application published January 22, 1992 under Publication No. 0467185 A2. In that publication, the oligomers of the present invention were described as having utility in the diagnosis and/or treatment of AIDS and AIDS related complex.
SUMMARY OF THE INVENTION The invention herein disclosed relates to a method of inhibiting the activity of inositol 1,4,5-triphosphate (IP3) by occupying the receptor sites specific to IP3 with a compound of the formula:
wherein n is a whole number selected from the range of 5 to ,fl 20 inclusive and the pharmaceutically acceptable salts thereof.
DETAILED DESCRIPTION OF THE INVENTION Inositol 1,4,5-triphosphate (IP3) is a naturally
15 occurring and active component of animal physiology. It is formed intracellularly upon the activation of cell-surface receptors linked to the enzyme phospholipase C. Once generated in sufficient quantities, IP3 acts to stimulate the release of calcium ions from storage organelles within
20 the cell. In this role IP3 is characterized as a "second messenger". Depending upon the type of cell, the calcium released by IP3 functions to stimulate a variety of physiologic processes such as smooth muscle contraction, histamine secretion and the hyperpolarization of nerve
25 cells. Any compound or agent that can promote or interfere with the function of IP3, will promote or interfere with the generation of calcium ions and thereby elicit predictable pharmacological effects.
30 The process by which IP3 releases calcium ions begins with the binding of IP3 to a specific receptor protein located on an intracellular calcium storage compartment located typically on the endoplasmic reticulum. This receptor protein has been cloned and has been shown to form
35 a calcium "channel" with unique structural properties when bound to IP3. Therefore, when IP3 binds with its receptor, a calcium channel is opened causing the release of calcium stored in the cell's endoplasmic reticulum. In turn, the
released calcium will elicit the appropriate cellular response.
Heretofore, the only verified potent antagonist of the IP3 receptor vas heparin, a complex glycosaminoglycan. The diamino benzenedisulfonic acid oligomers of this invention also appear to antagonize the effects of IP3 by competing for the receptor site. In most cases, these compounds are more effective than heparin and demonstrate fewer secondary effects. In addition to providing utility as laboratory
"tools" in evaluating the therapeutic potential of other IP3 receptor antagonists, the oligomers of this invention would also be administered to modulate IP3-induced calcium release and have a salutary effect on any number of disorders that are caused or exacerbated by an inordinately productive IP3 second messenger pathway.
EXPERIMENTALS Measurement of IP3 Binding Cerebella from male Sprague-Dawley rats (200 g) were homogenized in 30 volumes of ice-cold buffer A (50 mM Tris, 1 mM dithiothreitol, 1 mM EDTA, pH 7.7 with HC1) with a polytron (setting 9 for 10 seconds). The tissue is then washed twice by centrifugation (20,000 x g, 15 minutes; Sorvall 28-S., SS-34 rotor) and resuspended in 30 volumes of ice-cold buffer A.
For the binding assays, 1.5 ml eppendorf tubes containing 50 μl of test compound (made up as a lOx stock in water) or water, 50 μl [3H] IP3 (17Ci/mmol; Dupont-NEN; usually made as a 25 nM (lOx) stock solution in buffer), and 350 μl of buffer B (buffer A with pH adjusted to 8.4) on ice. Tubes for non-specific binding also contained 50 μl of non-radioactive IP3 (100 μM stock (lOx); final concentration 10 μM) , with an appropriate reduction in the volume of buffer B. Reactions were initiated by the addition of 50 μl tissue to make the final volume 500 μl.
followed by vortex mixing. Samples were incubated on ice for 10 minutes and then were centrifuged (14,000 x g) in a microfuge (Eppendorf model 5415) for 5 minutes followed by aspiration of the supernatant fraction. The tissue pellets were solubilized overnight in 100 μl of Protosol (Dupont- NEN). After solubilization, 73 μl of glacial acetic acid were added to decrease chemiluminescence, and the mixture was transferred to scintillation vials. To these vials was added 7 ml of Ecoscint-A (National Diagnostics) and the radioactivity determined by liquid scintillation spectrophotometry.
Specific binding was defined as the difference between total binding (radioactivity in the absence of test compound and cold IP3) and non-specific binding
(radioactivity in the absence of test compound but in the presence of cold IP3). This number was taken as 100% specific binding. Data points obtained with the test compounds were fit by a computer program (GraphPad-InPlot) to determine their inhibitory potency. The inhibitory potencies of the test compounds were expressed as the concentration of compound that produces 50% inhibition of specific binding (the IC50 value).
The binding data are presented in Table 1 and demonstrate that compounds within the scope of the present invention effectively compete for [3H] IP3 binding sites in rat cerebellar membranes. The compound identified as MDL 102,869 was the most potent competitor for binding with an IC50 of 50 nM, whereas low molecular weight heparin (5100 MW) had an IC50 of 74 nM. MDL 102,869 is the compound according to the claimed invention wherein n = 15.
The potency for binding also seems to correlate with the ability to antagonize IP3~induced calcium ion release. Thus, 1 and 3 μM of MDL 102,869 inhibited calcium ion release by 42 and 100%, respectively (see Fig. 1), whereas
10 μM of heparin inhibited release by 90%. MDL 101,828, which had an IC50 binding of 104 nM, inhibited IP3~induced calcium ion release by 72% at 3 micro moles. MDL 101,828 is the compound according to the claimed invention wherein n = 9.
TABLE 1
The tracing of Fig. 1, dramatically illustrates the IP3 inhibition data set forth in the third column of Table 1.
The y-axis represents the concentration of free calcium ions in arbitrary units. The tracing shows that two successive additions of 0.1 μM of IP3 stimulated similar amounts of calcium ion release from cerebellar microsomes. The addition of 1 μM of MDL 102,869 stimulated a small increase of calcium ion for unknown reasons. In the presence of 102,869, however, calcium ion release stimulated by 0.1 μM of IP3 was inhibited by 42%. This inhibition was overcome by the addition of 1 μM of IP3, consistent with competitive antagonism by MDL 102,869.
WHAT IS CLAIMED IS:
1. A method of inhibiting the activity of inositol 1,4,5-trisphosphate by occupying a receptor site specific to inositol 1,4,5-trisphosphate with a compound of the formula:
wherein n is a whole number within the range of 5-20 and the pharmaceutically acceptable salts thereof.
2. The oligomer of claim 1 wherein n is 9.
3. The oligomer of claim 1 wherein n is 15.
Claims (3)
1. A method of inhibiting the activity of inositol 5 1,4,5-trisphosphate by occupying a receptor site specific to inositol 1,4,5-trisphosphate with a compound of the formula:
wherein n is a whole number within the range of 5-20 and the pharmaceutically acceptable salts thereof.
2. The method of claim 1 wherein n is 9.
3. The method of claim 1 wherein n is 15.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US95239392A | 1992-09-28 | 1992-09-28 | |
PCT/US1993/008168 WO1994007507A1 (en) | 1992-09-28 | 1993-08-30 | Method for antagonizing inositol 1,4,5-triphosphate |
US952393 | 2007-12-07 |
Publications (2)
Publication Number | Publication Date |
---|---|
AU4841793A true AU4841793A (en) | 1994-04-26 |
AU670962B2 AU670962B2 (en) | 1996-08-08 |
Family
ID=25492868
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
AU48417/93A Ceased AU670962B2 (en) | 1992-09-28 | 1993-08-30 | Method for antagonizing inositol 1,4,5-triphosphate |
Country Status (13)
Country | Link |
---|---|
EP (1) | EP0661982A1 (en) |
JP (1) | JPH08502068A (en) |
KR (1) | KR950703347A (en) |
AU (1) | AU670962B2 (en) |
CA (1) | CA2145681A1 (en) |
HU (1) | HUT70192A (en) |
IL (1) | IL107066A (en) |
MX (1) | MX9305933A (en) |
NO (1) | NO951163D0 (en) |
NZ (1) | NZ256178A (en) |
TW (1) | TW260663B (en) |
WO (1) | WO1994007507A1 (en) |
ZA (1) | ZA936978B (en) |
Families Citing this family (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU2978995A (en) * | 1994-06-30 | 1996-01-25 | Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. | Methods for treating cell proliferative disorders by modulating signal transduction |
EP0809492A4 (en) * | 1995-02-17 | 2007-01-24 | Smithkline Beecham Corp | Il-8 receptor antagonists |
US5780483A (en) * | 1995-02-17 | 1998-07-14 | Smithkline Beecham Corporation | IL-8 receptor antagonists |
US6083920A (en) * | 1995-12-21 | 2000-07-04 | Ayurcore, Inc. | Compositions for modulating intracellular inositol trisphosphate concentration |
US6005008A (en) * | 1996-02-16 | 1999-12-21 | Smithkline Beecham Corporation | IL-8 receptor antagonists |
US6262113B1 (en) | 1996-03-20 | 2001-07-17 | Smithkline Beecham Corporation | IL-8 receptor antagonists |
US6211373B1 (en) | 1996-03-20 | 2001-04-03 | Smithkline Beecham Corporation | Phenyl urea antagonists of the IL-8 receptor |
HUP9903922A3 (en) | 1996-06-27 | 2001-12-28 | Smithkline Beecham Corp | Il-8 receptor antagonists |
KR20000022274A (en) * | 1996-06-27 | 2000-04-25 | 스튜어트 알. 수터, 스티븐 베네티아너, 피터 존 기딩스 | Il-8 receptor antagonist |
WO2002038140A2 (en) * | 2000-11-09 | 2002-05-16 | Contrimmune Biotechnology Inc | Therapeutic uses for ip3 receptor-mediated calcium channel modulators |
EP2750767A4 (en) | 2011-09-02 | 2015-10-14 | Univ Columbia | CaMKII, IP3R, CALCINEURIN, P38 AND MK2/3 INHIBITORS TO TREAT METABOLIC DISTURBANCES OF OBESITY |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB781479A (en) * | 1953-08-27 | 1957-08-21 | Ciba Ltd | New polyureas and process for making them |
US5276182A (en) * | 1990-07-09 | 1994-01-04 | The Dow Chemical Company | Process for preparing polyurea oligomers |
-
1993
- 1993-08-30 AU AU48417/93A patent/AU670962B2/en not_active Ceased
- 1993-08-30 CA CA002145681A patent/CA2145681A1/en not_active Abandoned
- 1993-08-30 WO PCT/US1993/008168 patent/WO1994007507A1/en not_active Application Discontinuation
- 1993-08-30 EP EP93921252A patent/EP0661982A1/en not_active Ceased
- 1993-08-30 KR KR1019950701166A patent/KR950703347A/en not_active Application Discontinuation
- 1993-08-30 NZ NZ256178A patent/NZ256178A/en unknown
- 1993-08-30 HU HU9500891A patent/HUT70192A/en unknown
- 1993-08-30 JP JP6509054A patent/JPH08502068A/en active Pending
- 1993-09-21 ZA ZA936978A patent/ZA936978B/en unknown
- 1993-09-22 TW TW082107787A patent/TW260663B/zh active
- 1993-09-22 IL IL107066A patent/IL107066A/en not_active IP Right Cessation
- 1993-09-27 MX MX9305933A patent/MX9305933A/en unknown
-
1995
- 1995-03-27 NO NO951163A patent/NO951163D0/en unknown
Also Published As
Publication number | Publication date |
---|---|
CA2145681A1 (en) | 1994-04-14 |
TW260663B (en) | 1995-10-21 |
NZ256178A (en) | 1997-03-24 |
WO1994007507A1 (en) | 1994-04-14 |
IL107066A0 (en) | 1993-12-28 |
NO951163L (en) | 1995-03-27 |
HU9500891D0 (en) | 1995-05-29 |
KR950703347A (en) | 1995-09-20 |
EP0661982A1 (en) | 1995-07-12 |
AU670962B2 (en) | 1996-08-08 |
MX9305933A (en) | 1994-04-29 |
IL107066A (en) | 1998-01-04 |
HUT70192A (en) | 1995-09-28 |
NO951163D0 (en) | 1995-03-27 |
ZA936978B (en) | 1994-04-18 |
JPH08502068A (en) | 1996-03-05 |
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