METHODS OF TREATING RENAL DISEASE WITH A GASTRIN RELEASING PEPTIDE RECEPTOR ANTAGONIST

Abstract: Invented is a method of treating renal disease, preferably chronic renal disease, in a mammal, including human, which comprises administering to such mammal an effective amount of a Gastrin Releasing Peptide Receptor antagonist.

Description: METHODS OF TREATING RENAL DISEASE WITH A GASTRIN RELEASING PEPTIDE RECEPTOR ANTAGONIST This invention relates to a method of treating renal disease, preferably chronic renal disease, in a mammal, including a human, in need thereof which comprises administering to such mammal an effective amount of a Gastrin Releasing Peptide Receptor (GRP-R) antagonist. Background of the Invention Renal disease is a multifactorial process which involves cell proliferation and matrix deposition. These are followed by glomerulosclerosis and tubular as well as interstitial fibrosis. Current therapies for renal disease and particularly chronic renal disease (treatment of underlying disease, control of hypertension, protein restriction or ACE inhibitors) have demonstrated limited efficacy. Thus, there is a need in the art for a new method of treating renal disease and particularly chronic renal disease. Gastrin releasing peptide (GRP) is a 27 amino acid peptide having the following sequence in humans: Val-Pro-Leu-Pro-Ala-Gly-Gly-Gly-Thr-Val-Leu-Thr-Lys-Met-Tyr-Pro-Arg-Gly- Asn-His-T -Ala-Val-Gly-His-Leu-Met-NH2- GRP is indicated as an autocrine factor in several types of cancer cells. Dietrich, J. B. Cell Mol. Bio. 40: 731 -746, (1994) and Cuttitta, F. et al.. Nature 316: 823-826, (1985). Specifically, GRP has been shown to stimulate proliferation of lung (Cuttitta, F. et al., Nature 316: 823- 826, (1985)), breast (Nelson, J. et al., Br. J. Cancer 63: 933-936, (1991)) and prostate (Bologna, M. et al., Cancer 63: 1714-1720 (1989)) carcinoma cells. Several lung and prostate cancer cells which proliferate in response to GRP, were found to be inhibited by treatment with GRP antibodies (Maruno, K. et al., Life Sci. 51- 267-71 (1993)) and GRP receptor antagonists (Moody, T. W., Life Sci. 56: 521-529 (1995)). In addition, the growth of prostate and breast cancer cell xenografts in a nude mouse host were found to be inhibited by treatment with GRP receptor antagonists (Pinski J., et al. Cancer Lett. 71: 189-196 (1993) and Shirahige Y., Biomed. Pharmacother. 4£: 465-472 (1994)) and GRP antibodies (Shimoda, J., Japanese Urol. J. 48: 465-472 (1994)). Presently, antagonists of the GRP receptor are not known as having utility in treating renal disease and particularly not in treating chronic renal disease. Summary of the Invention This invention relates to a method of treating renal disease, preferably chronic renal disease, in a mammal, including a human, in need thereof which comprises administering to such mammal an effective amount of a Gastrin Releasing Peptide Receptor (GRP-R) antagonist. Brief Description of the Drawings FIG. 1 (top panel) depicts the competition of [1251] aGRP14-27 binding to rat mesangial cells by unlabeled GRP and NMB; FIG. 1 (bottom panel) depicts scatchard analysis of [1251] aGRP 14-27 binding to rat mesangial cells; FIG. 2 depicts the effect of GRP on (Ca++)i mobilization in rat mesangial cells; FIG. 3 depicts the effect of GRP on mesangial cell proliferation; and FIG. 4 depicts the effect of GRP on TGF βl, fibronectin and osteopontin mRNA in mesangial cells. Detailed Description of the Invention All publications, including but not limited to patents and patent applications, cited in this specification are herein incorporated by reference as though fully set forth. By the term "treating" as used herein is meant prophylactic or therapeutic therapy. By the term "Gastrin Releasing Peptide Receptor antagonists" and derivatives thereof, as used herein is meant an antibody, a peptide or a nonpeptide chemical compound which exhibits pharmaceutical activity by antagonizing the GRP Receptor. Persons skilled in the art can readily determine if an antibody, a peptide or a nonpeptide chemical compound is a Gastrin Releasing Peptide Receptor antagonists by known methods. All such agents are included within the scope of the invention. It has now been discovered that compounds which are antagonists of the Gastrin Releasing Peptide (GRP) Receptor are useful in treating renal disease, preferably chronic renal disease. This discovery was demonstrated by the Mesangial Cell Membrane Preparation and [-^I] Bombesin Binding assay described below. To perform the experiments Mesangial cell membranes were prepared following the procedure of Albrightson et al. Mol. Pharmacol.. 47: 1156-1163 (1995). Increasing concentrations of [12 τj Bombesin were added to tubes containing mesangial cell membranes (10-20 ug/tube) and incubated for 30 min. at 30°C. Non-specific binding was measured in the presence of 1 uM unlabeled bombesin. Bound and free ligands were separated by filtration through Whatman GF/C filters presoaked in 0.1 % BS A. The filters were washed three times with buffer and counted in a gamma counter with an efficiency of 75%. Specific binding which was between 85-95% was calculated by subtracting non-specific binding from total binding. Competition binding experiments were performed by adding increasing concentrations of unlabeled ligands to membranes in the presence of indicated concentration of [**^I] Bombesin. Measurement of Intracellular Calcium Intracellular calcium mobilization in response to GRP was determined by measuring the fluorescence of fura-2-loaded mesangial cells with a dual-wave length spectrofluorometer (University of Pennsylvania Biomedical Instruments Group) following the procedure of Albrightson et al. (1995). Cell Proliferation Cell proliferation was measured as described in Albrightson et al., J. Pharmacol. Exp. Ther. 263: 404-412, (1992). Mesangial cells in 24 well plates were exposed to increasing concentrations of GRP for 16-24 hr and then challenged with [3.H] thymidine for 4 hr. At the end of the incubation, the cells were washed and precipitated with TCA and counted. Isolation of RNA and Northern Blot Analysis Total RNA was isolated from mesangial cells following the procedure of Chomczynski et al., Anal. Biochem. 162: 156-159, (1987) with modifications (Albrightson et al. 1995). Northern blot analysis was done as explained (Albrightson et al., 1995). Results Addition of increasing concentrations of unlabeled GRP and neuromedin B to mesangial cell membranes displaced [-^I] Bombesin binding in a concentration dependent manner. Neuromedin B was ~ 100-fold weaker than GRP in competing for the binding sites (Figure 1, top panel). Addition of increasing concentrations of [125i] Bombesin resulted in specific, saturable and high affinity binding. The apparent dissociation constant (K^j) and maximum binding (B max ) were 84 pM and 100 fmol/mg protein, respectively (Figure 1, bottom panel). Addition of increasing concentrations of GRP to fura-2-loaded mesangial cells resulted in a concentration-dependent increase in the release of intracellular calcium with approximate EC50 va lue °f 3-10 nM (Figure 2). Also, exposure of mesangial cells to increasing concentrations of GRP resulted in concentration- dependent increase in [*-Η] thymidine incorporation which is indicative of cell proliferation (Figure 3). In addition, GRP caused an increase in TGFβl, fibronectin and osteopontin mRNA levels which is indicative of matrix formation (Figure 4). The results of the above experiments clearly demonstrates the therapeutic utility of Gastrin Releasing Peptide Receptor antagonist on treating renal disease, particularly chronic renal disease. This invention discloses Gastrin Releasing Peptide Receptor antagonist and pharmaceutically acceptable salts or hydrates or solvates thereof as being useful for treating renal disease, preferably chronic renal disease, in mammals, including humans. A Gastrin Releasing Peptide Receptor antagonist or a pharmaceutically acceptable salt or hydrate or solvate thereof can be administered to a subject in a conventional dosage form prepared by combining a Gastrin Releasing Peptide Receptor antagonist or a pharmaceutically acceptable salt or hydrate or solvate thereof, with a conventional pharmaceutically acceptable carrier or diluent according to known techniques. It will be recognized by one of skill in the art that the form and character of the pharmaceutically acceptable carrier or diluent is dictated by the amount of active ingredient with which it is to be combined, the route of administration and other well-known variables. A Gastrin Releasing Peptide Receptor antagonist or a pharmaceutically acceptable salt or hydrate or solvate thereof is administered to a mammal, including a human, in an amount sufficient to prevent or alleviate renal disease, preferably chronic renal disease. The route of administration of the Gastrin Releasing Peptide Receptor antagonist is not critical but is usually oral or parenteral, preferably oral. The term parenteral as used herein includes intravenous, intramuscular, subcutaneous, intranasal, intrarectal, transdermal, intravaginal or intraperitoneal administration. The subcutaneous and intramuscular forms of parenteral administration are generally preferred. The daily parenteral dosage regimen will preferably be from about 0.01 mg kg to about 50 mg/kg of total body weight, most preferably, from about 0.1 mg/kg to about 5 mg/kg. Preferably, each parenteral dosage unit will contain the active ingredient in an amount of from about 0.5 mg to about 150 mg. The Gastrin Releasing Peptide Receptor antagonist which are active when given orally can be formulated as liquids, for example, syrups, suspensions or emulsions, tablets, capsules and lozenges. A liquid formulation will generally consist of a suspension or solution of the compound or pharmaceutically acceptable salt in a suitable liquid carrier(s) for example, ethanol, glycerine, non-aqueous solvent, for example polyethylene glycol, oils, or water with a suspending agent, preservative, flavoring or coloring agent. A composition in the form of a tablet can be prepared using any suitable pharmaceutical carrier(s) routinely used for preparing solid formulations. Examples of such carriers include magnesium stearate, starch, lactose, sucrose and cellulose. A composition in the form of a capsule can be prepared using routine encapsulation procedures. For example, pellets containing the active ingredient can be prepared using standard carriers and then filled into a hard gelatin capsule; alternatively, a dispersion or suspension can be prepared using any suitable pharmaceutical carrier(s), for example aqueous gums, celluloses, silicates or oils and the dispersion or suspension then filled into a soft gelatin capsule. The daily oral dosage regimen will preferably be from about 0.01 mg/kg to about 50 mg/kg of total body weight. Preferably each oral dosage unit will contain the active ingredient in an amount of from about 0.5 mg to about 150 mg. While it is possible for an activate ingredient to be administered alone, it is preferable to present it as a pharmaceutical formulation. It will be recognized by one of skill in the art that the optimal quantity and spacing of individual dosages of a Gastrin Releasing Peptide Receptor antagonist or a pharmaceutically acceptable salt or hydrate or solvate thereof will be determined by the nature and extent of the exact condition being treated, the form, route and site of administration, and the particular patient being treated, and that such optimums can be determined by conventional techniques. It will also be appreciated by one of skill in the art that the optimal course of treatment, i.e., the number of doses of a Gastrin Releasing Peptide Receptor antagonist or a pharmaceutically acceptable salt or hydrate or solvate thereof given per day and duration of therapy, can be ascertained by those skilled in the art using conventional course of treatment determination tests. The method of this invention of treating renal disease, preferably chronic renal disease, in mammals, including humans, comprises administering to a subject in need of such treatment an effective amount of a pharmaceutically active compound of the present invention. The invention also provides for the use of a Gastrin Releasing Peptide Receptor antagonist or a pharmaceutically acceptable salt or hydrate or solvate thereof in the manufacture of a medicament for use in treating renal disease, preferably chronic renal disease, in mammals, including humans. The invention also provides for a pharmaceutical composition for use in the treatment of renal disease, preferably chronic renal disease, which comprises a Gastrin Releasing Peptide Receptor antagonist or a pharmaceutically acceptable salt or hydrate or solvate thereof. The invention also provides for a process for preparing a pharmaceutical composition containing a Gastrin Releasing Peptide Receptor antagonist or a pharmaceutically acceptable salt or hydrate or solvate thereof which comprises bringing the Gastrin Releasing Peptide Receptor antagonist or a pharmaceutically acceptable salt or hydrate or solvate thereof into association with the pharmaceutically acceptable carrier or diluent. No unacceptable toxicological effects are expected when compounds of the invention are administered in accordance with the present invention. In addition, the compounds of the present invention can be co-administered with further active ingredients, such as compounds known to treat renal disease and particularly chronic renal disease. Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. The following examples are, therefore, to be construed as merely illustrative and not a limitation of the scope of the present invention in any way. Example 1 - Capsule Composition A oral dosage form for administering a Gastrin Releasing Peptide Receptor antagonist is produced by filing a standard two piece hard gelatin capsule with the ingredients in the proportions shown in Table I, below. Table I INGREDIENTS AMOUNTS GRP Releasing Peptide antagonist 25 mg Lactose 55 mg Talc 16 mg Magnesium Stearate 4 mg Example 2 - Injectable Parenteral Composition An injectable form for administering a Gastrin Releasing Peptide Receptor antagonist is produced by stirring 1.5% by weight of a GRP Releasing Peptide antagonist in 10% by volume propylene glycol in water. Example 3 - Tablet Composition The sucrose, calcium sulfate dihydrate and a Gastrin Releasing Peptide Receptor antagonist shown in Table II below, are mixed and granulated in the proportions shown with a 10% gelatin solution. The wet granules are screened, dried, mixed with the starch, talc and stearic acid, screened and compressed into a tablet. Table II INGREDIENTS AMOUNTS GRP Releasing Peptide antagonist 20 mg calcium sulfate dihydrate 30 mg sucrose 4 mg starch 2 mg talc 1 mg stearic acid 0.5 mg While the above descriptions and examples fully describe the invention and the preferred embodiments thereof, it is understood that the invention is not limited to the particular disclosed embodiments coming within the scope of the following claims.

Claims: What is claimed is: 1. A method of treating renal disease in a mammal which comprises administering to such mammal an effective amount of a Gastrin Releasing Peptide Receptor antagonist. 2. A method of claim 1 in which the mammal is a human. 3. The method of claim 2 wherein the Gastrin Releasing Peptide Receptor antagonist is administered orally. 4. The method of claim 3 wherein from about 0.01 mg/kg to about 20 mg/kg of the Gastrin Releasing Peptide Receptor antagonist is administered per day. 5. The method of claim 2 wherein the Gastrin Releasing Peptide Receptor antagonist is administered parenterally. 6. The method of claim 5 wherein from about 0.01 mg to about 20 mg/kg of the Gastrin Releasing Peptide Receptor antagonist is administered per day. 7. A method of treating chronic renal disease in a mammal which comprises administering to such mammal an effective amount of a Gastrin Releasing Peptide Receptor antagonist. 8. A method of claim 7 in which the mammal is a human. 9. The method of claim 8 wherein the Gastrin Releasing Peptide Receptor antagonist is administered orally. 10. The method of claim 9 wherein from about 0.01 mg/kg to about 20 mg/kg of the Gastrin Releasing Peptide Receptor antagonist is administered per day. 1 1. The method of claim 8 wherein the Gastrin Releasing Peptide Receptor antagonist is administered parenterally. 12. The method of claim 1 1 wherein from about 0.01 mg to about 20 mg/kg of the Gastrin Releasing Peptide Receptor antagonist is administered per day. 13. The use of a Gastrin Releasing Peptide Receptor antagonist in the manufacture of a medicament for use in treating renal disease in a mammal. 14. A use according to claim 13 in which the mammal is a human. 15. A use according to claim 14 wherein the Gastrin Releasing Peptide Receptor antagonist is administered orally. 16. A use according to claim 15 wherein from about 0.01 mg/kg to about 20 mg/kg of the Gastrin Releasing Peptide Receptor antagonist is administered per day. 17. A use according to claim 14 wherein the Gastrin Releasing Peptide Receptor antagonist is administered parenterally. 18. A use according to claim 17 wherein from about 0.01 mg to about 20 mg/kg of the Gastrin Releasing Peptide Receptor antagonist is administered per day. 19. The use of a Gastrin Releasing Peptide Receptor antagonist in the manufacture of a medicament for use in treating chronic renal disease in a mammal. 20. A use according to claim 19 in which the mammal is a human. 21. A use according to claim 20 wherein the Gastrin Releasing Peptide Receptor antagonist is administered orally. 22. A use according to claim 21 wherein from about 0.01 mg/kg to about 20 mg/kg of the Gastrin Releasing Peptide Receptor antagonist is administered per day. 23. A use according to claim 20 wherein the Gastrin Releasing Peptide Receptor antagonist is administered parenterally. 24. A use according to claim 23 wherein from about 0.01 mg to about 20 mg/kg of the Gastrin Releasing Peptide Receptor antagonist is administered per day. 25. A pharmaceutical composition for use in treating renal disease in a mammal comprising a Gastrin Releasing Peptide Receptor antagonist and a pharmaceutically acceptable carrier. 26. A composition according to claim 25 in which the mammal is a human. 27. A composition according to claim 26 wherein the Gastrin Releasing Peptide Receptor antagonist is administered orally. 28. A composition according to claim 27 wherein from about 0.01 mg/kg to about 20 mg/kg of the Gastrin Releasing Peptide Receptor antagonist is administered per day. 29. A composition according to claim 26 wherein the Gastrin Releasing Peptide Receptor antagonist is administered parenterally. 30. A composition according to claim 29 wherein from about 0.01 mg to about 20 mg/kg of the Gastrin Releasing Peptide Receptor antagonist is administered per day. 31. A pharmaceutical composition for use in treating chronic renal disease in a mammal comprising a Gastrin Releasing Peptide Receptor antagonist and a pharmaceutically acceptable carrier. 32. A composition according to claim 31 in which the mammal is a human. 33. A composition according to claim 32 wherein the Gastrin Releasing Peptide Receptor antagonist is administered orally. 34. A composition according to claim 33 wherein from about 0.01 mg/kg to about 20 mg/kg of the Gastrin Releasing Peptide Receptor antagonist is administered per day. 35. A composition according to claim 32 wherein the Gastrin Releasing Peptide Receptor antagonist is administered parenterally. 36. A composition according to claim 35 wherein from about 0.01 mg to about 20 mg/kg of the Gastrin Releasing Peptide Receptor antagonist is administered per day.

Inventors: NAMBI PONNAL, NAMBI, PONNAL
Applicants: NAMBI PONNAL, SMITHKLINE BEECHAM CORP, NAMBI, PONNAL, SMITHKLINE BEECHAM CORPORATION
Application year: 1997
Main classification: A61K38
IPC codes: A61K38, A61K39, A61K45, A61P13, A61P43